Announcing the launch of the new Ludger Glycan standards webpages

Oxford – March 2021

Ludger - Glycan Standards Webpages

We are very excited at Ludger to launch our newly designed Glycan Standards webpages with immediate effect. Ludger has a range of system suitability standards, process controls and reference standards for different applications including the analysis of sialic acids, monosaccharides, glycan profiling and characterisation. We provide a variety of unlabelled N-glycans (N), O-glycans (O), oligosaccharides, glycoproteins / glycopeptides and glycan library standards. Furthermore, our catalogue of standards also includes a large range of N- and O- glycan standards fluorescently labelled with 2-aminobenzamide (2-AB), 2-AA (2-aminobenzoic acid), procainamide, 8-aminopyrene-1,3,6-trisulfonic acid (APTS) and permethylated glycan standards. We also provide quantitative glycan and glycopeptide standards under the brand name Ludger BioQuant. Please visit our standards webpages for more information.

Ludger glycan and glycopeptide standards can be used for the following applications:
(i) Process controls for release, labelling, exoglycosidase sequencing and analysis
(ii) System suitability testing for MS, (U)HPLC and CE
(iii) Reference standards (e.g., GU (glucose unit) calibration for (U)HPLC)
(iv) Structure identification
(v) Quantitation

The new webpages have been designed with an intuitive layout for ease of navigation and the menu options are designed to guide you to the most appropriate standard of your choice for your glycan analysis workflows.

We have incorporated valuable information regarding each standard by including product specifications, glycan cartoon depicting the glycan structure and the mass. Each standard page also has links to their associated the generic product guides, specification sheets and safety data sheets to assist you with glycan profiling and characterisation.

For more information, please visit www.ludger.com/products/glycan-standards.
If you have any questions or to request a quotation, please contact us at info@ludger.com. We sincerely hope you enjoy navigating through our new standards webpages and if you have any suggestions or feedback please E-mail us.






Product Discontinuation Notice

Oxford – April 2021

Ludger logo

Ludger will discontinue the following LudgerClean products listed below as of 1st June 2021 or until stocks last.

Serial number Ludger catalogue number Product description
1 LC-PERMET-96 LudgerClean Pre-Permethylation clean-up plate
2 LC-PROC-96 LudgerClean Procainamide clean-up plate


The discontinuance of these products is due to a raw material discontinuation from our supplier. During the replacement process, we have evaluated and performed the necessary comparability studies between the old and the new Ludger clean range plates.

We appreciate and value our business relationship with you and look forward to your continued interest in our products. While we regret any inconvenience this announcement may cause, we are pleased to announce that we have alternate products that will replace these plates. They will use the same catalogue numbers and will provide the same or better performance in our glycan clean up workflows in comparison to the discontinued products.

Please contact info@ludger.com if you have additional questions or interested in the validation study report and/or to discuss how we can support you through this conversion.






GLYCO 2030 - A Roadmap for Glycoscience in Europe

Oxford – March 2021

Ludger - Glyco 2030 - A Roadmap for Glycoscience in Europe

Ludger is very happy to have been a part of the glycoscience community in Europe and for publishing the article GLYCO 2030-A Roadmap for Glycoscience in Europe in collaboration with CarboMet.

The production of this glycoscience roadmap was made possible thanks to support from the European Commission and the Glycoscience community in Europe. The journey started with a workshop in 2016 called “Metrology of Carbohydrates” which was attended by glycoscientists from industry and academia. Through a series of interactive group discussions, the four CarboMet bioindustry sectors (BIS) emerged along with the three Enabling Technology Areas (ETAs) as areas which showed enormous innovation potential. CarboMet was established to ensure the ideas generated in the workshop was realized through a series of focused workshop in each BIS, publication of policy papers and with the summation of the glycoscience roadmap. The aim of the roadmap is to highlight the challenges and opportunities in European research and innovation and where glycoscience can contribute to the advancement in the four BIS.

You can view the GLYCO2030 roadmap by clicking here. For any questions, suggestions, feedback or comments, please contact us at info@ludger.com







Publication in PLOS Neglected Tropical Diseases: Tsetse salivary glycoproteins are modified with paucimannosidic N-glycans, are recognised by C-type lectins and bind to trypanosomes

Oxford – March 2021

Ludger PLOS Neglected Tropical Diseases Publication - Tsetse Fly
Photo credit: Oregon State University © 2011

Successful collaboration between Ludger and The Liverpool School of Tropical Medicine, resulted in publishing an article in PLOS Neglected Tropical Diseases titled “Tsetse salivary glycoproteins are modified with paucimannosidic N-glycans, are recognised by C-type lectins and bind to trypanosomes.”

African sleeping sickness is caused by Trypanosoma brucei, a parasite transmitted by the bite of a tsetse fly. Most research has focused on the salivary proteins, while glycans that modify them remain unexplored. Insect salivary glycans may affect how the saliva is recognized by the host, possibly playing a role during pathogen transmission. This is the first study of the salivary glycans of tsetse fly Glossina morsitans providing detailed information about their structures and relative abundances. This study shows that tsetse fly glycoproteins are mainly modified by simple N-glycans with short mannose modifications, which are recognised by mammalian C-type lectins (mannose receptor and DC-SIGN). This information provokes interesting questions as to the role of these glycoproteins in the successful establishment of infection by this parasite.

Ludger’s contribution to this study included:

  • Help with the study design and strategy
  • Release of N- and O-glycans from tsetse fly saliva
  • N- and O-glycan profiling and characterisation using orthogonal techniques such as UHPLC, LC-ESI-MS and exoglycosidase digestions (see Figure 1 for data on salivary N-glycans from Tsetse flies, before and after digestion with exoglycosidases)
  • Supporting the visual reporting of structures and nomenclature of glycans identified in the study

Ludger PLOS Neglected Tropical Diseases - Tsetse Fly - Figure 1
Figure 1: Profile of salivary N-glycans from teneral (young, unfed) flies, before and after digestion with exoglycosidases. Aliquots of the total PNGase F-released 2-AB-labeled N-glycan pool were either undigested (i) or incubated with a range of exoglycosidases (ii-iv). (i) Undig, before digestion; (ii) GUH, Streptococcus pneumoniae in E. coli β-N-acetylglucosaminidase; (iii) JBM, Jack bean α-Mannosidase; (iv) bkF, Bovine kidney α-fucosidase. Following digestion, the products were analyzed by HILIC-UHPLC.

To find out how to utilise enzymes in glycan characterisation visit our Exoglycosidase enzyme page. Visit our Procainamide webpage for more information on how to characterise glycans using LC-MS. And for more information about this article visit our Publications webpage.






Ludger Glycoanalysis-Tools: labelling kits containing the non-toxic reducing agent 2-picoline borane (2PB)

Oxford – March 2021

Fluorescent labelling methods for glycoanalysis of therapeutic glycoproteins are well established and aid subsequent separation and quantitation of glycans using a range of techniques. These include high performance liquid chromatography (HPLC and UHPLC), capillary electrophoresis (CE), and mass spectrometry or a combination thereof. Ludger provides a range of labelling kits to fluorescently label N- and O-glycans, that have been released enzymatically or chemically. These include 2-aminobenzamide (2AB) labelling, procainamide (PROC) labelling and 2-aminobenzoic acid (2AA) labelling kits.

Ludger 2PB - Figure 1
Figure 1: Overlay of 8 human IgG N-glycan samples analysed using a HILIC UHPLC column: four samples labelled using the procainamide labelling kit with 2PB as the reductant and four samples labelled using the procainamide labelling kit with sodium cyanoborohydride as the reductant

Many commercially available 2AB, PROC and 2AA labelling kits use sodium cyanoborohydride as the reducing agent during glycan labelling. Although this reductant is a gold standard it is toxic and therefore the best practice is to perform the labelling in a fume cupboard for safe handling. To provide a safer, less toxic option that can be used on a laboratory bench without the need to use a fumehood, Ludger offers labelling kits that use 2-picoline borane (2PB), which is a significantly safer reductant.

Features and benefits of LudgerTag™ 2PB kits
The labelling kits containing 2PB as the reductant are equally efficient with respect to glycan labelling performance as the traditional sodium cyanoborohydride kits (See Figure 1, a typical chromatogram overlay of PROC labelled human IgG N-glycans).

Fully validated Ludger labelling kits
The 2PB containing kits have been validated following ICH Q2(R1) guidelines. Precision values are excellent, with coefficient of variation (CV’s) of less than 5% for peaks with relative % areas greater than 5% and with CVs of less than 8% for peaks with relative % areas less than 5%. The kits offer the equivalent labelling efficiency as the standard Ludger Tag 2AA, 2AB and PROC labelling kits containing sodium cyanoborohydride.

Ludger Workflow for N and O-glycan labelling

Improved safety
2PB has a lower toxicity compared to sodium cyanoborohydride and therefore is safer.

Ease of use
Each kit comprises three bottles or vials; one containing acetic acid and DMSO solution, the second vial has the reductant 2PB and the third vial has the dye (2AB, 2AA or PROC). As there are fewer vials involved in the make- up of the labelling solution, it makes the labelling process simpler and more efficient.

For more information on our labelling kits containing 2PB as the reductant please visit: our products page for Glycan Labelling and Derivatisation Kits. For enquiries or more information, please contact: info@ludger.com






Ludger Application Note: O-glycosylation Analysis

Oxford – February 2021

Ludger Application Note - O-glycosylation Analysis

O-glycosylation is known to have a critical impact on protein secretion and protective immunity against cancer, and maintenance of normal development and physiology. Moreover, a substantial portion of the protein-based pharmaceuticals on the market are glycoproteins where O-glycosylation has found to critically modulate the physiochemical properties of proteins, have a functional impact on their therapeutic potentials, and affect the safety profiles of such drugs. O-glycosylation is prevalent in many classes of therapeutic proteins including Erythropoietin (EPO), Follicle stimulating hormone (FSH), Etanercept, Granulocyte-colony stimulating factor (G-CSF) providing evidence of its critical involvement in drug performance and diseases.

Our application note provides the details on how to perform O-glycan analysis using Ludger’s glyco-analytical technology which includes:

  • O-glycan release: by using Ludger enzyme (E-G001) for release of unsubstituted Galβ1,3-GalNAcα disaccharides and/or Ludger Liberate Hydrazinolysis (LL-HYDRAZ-A2) and Orela (LL-ORELA-A2) kits for complete removal of O-glycans from glycoproteins or glycopeptides
  • O-glycan derivatisation
  • Use of system suitability standards

To enquire or place an order please contact: info@ludger.com

For more information and to view this and our other application notes, please visit our Support-Resources webpages.






Characterising Glycans from SARS-CoV-2 Specimen Samples and Vaccine Candidates using Ludger’s Custom Analytical Services

Oxford – January 2021

Ludger Glycan Analysis Services - SARS-CoV-2 Specimen samples

The current global situation with Coronavirus disease 2019 (COVID-19) has changed our daily lives and the way we interact.

The COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Understanding the N-glycosylation of the SARS-CoV-2 viral spike protein variations and its potential effects on the interaction with host immune system is essential in the vaccine development and for understanding the disease progression.

Ludger offers custom analytical services to suit your individual requirements. We have many years of expertise with analysing glycosylation (including N- and O- glycosylation) from variety of sample types and our laboratories are prepared to receive:

  • COVID-19 patient samples (e.g. plasma, tissues)
  • SARS-CoV-2 infected cell lines
  • Vaccine candidates
  • mAbs, glycoprotein hormones, Fc fusion proteins

For more information, please visit the Glycan Analysis Services pages.

If you have any questions relating to glycan analysis, or to request a Study Proposal and/or quotation, please contact us at info@ludger.com or contact Dr Radoslaw Kozak (Head of Glycoprofiling, Ludger) at rad.kozak@ludger.com






Business Development Assistant vacancy at Ludger

Oxford – January 2021

Ludger logo

Applications are invited for a Business Development (BD) Assistant position at Ludger. Located in Oxfordshire, we specialise in the development and manufacture of glycan characterisation technology and glycan analysis services to the biopharmaceutical industry. Our clients include top biotechnology companies and research organisations worldwide. Ludger is also expanding its client offering outside of the traditional glycobiology range which involves precision medicine research that will eventually develop into service/product offerings.

We currently have the requirement for a BD Assistant who will report directly to the BD Lead. The position will cover several aspects of BD with a focus on increasing new lead generation, client conversion, sales, and marketing.

Candidates must be eligible to live and work in the UK. A competitive salary will be offered, commensurate with qualifications and experience.

For more information on this role please view the Job Description
Application deadline: 12 Feb 2021
Please apply by emailing a covering letter and CV to: sophie.davey@ludger.com






Publication in Glycoconjugate Journal: A novel glycosidase plate-based assay for the quantification of galactosylation and sialylation on human IgG

Oxford – November 2020

Ludger Glycoconjugate Journal Publication - Figure 1
Fig 1. The released N-glycans from IgG glycoproteins (a) without galactosidase treatment was compared with (b) galactosidase treatment on a HILIC-FLD-MSn platform after labelling the reducing end with procainamide

Ludger is pleased to announce the publication in the Glycoconjugate Journal of a scientific paper written by Osmond Rebello who was one of our Marie Sklodowska Curie early stage researchers on the EU Horizon 2020 funded project GlySign (www.glysign.eu) . This paper explores the use of a microtitre plate based assay for assessing the galactose levels present in protein G extracted IgG from patient blood samples and comparing the amounts detected with a higher end LC-MS technique. The plate based assay contains an exoglycosidase enzyme that was developed by our Innovate UK/BBSRC IBCatalyst funded collaborators Drs Lucy Crouch and David Bolam as part of the Glycoenzymes for Bioindustries project (www.glycoenzymes.com).

Immunoglobulin IgG circulating in people is a glycosylated protein which contains a single glycan per Fc region. This glycan lends itself to bioactivities within people with the level of galactosylation being linked to inflammation mechanisms of the immune system. Lower levels of IgG galactosylation are seen in patients with diseases of inflammation such as rheumatoid arthritis and inflammatory bowel disease as well as in people with high levels of chronic inflammation and those of advanced age, known as “inflammaging”. Currently technique for measuring the galactose levels will involve high-end instrumentation such as UHPLC and mass spectrometry. This paper explored the use of a simple plate based assay for measuring galactose and compared this favourable to the results using UHPLC/HILIC. Once prepared galactose levels can be determined for hundreds of samples within minutes.

To find out how to utilise enzymes in glycan characterisation visit our Exoglycosidase enzyme page. Visit our Procainamide webpage for more information on how to characterise glycans using LC-MS. And for more information about this article visit our Publications webpage.






GlySign consortium video

Oxford – October 2020

GlySign consortium video

GlySign Early Stage Researchers prepared a video summarizing a three-year research training network, which Ludger was a part of. The network gathered two industrial partners (Ludger in the UK and Genos in Croatia) and an academic partner, Leiden University Medical Center (LUMC), in the Netherlands. The research focus was on glycomic markers and assay development for precision medicine covering a range of diseases including diabetes, prostate cancer and immune diseases. This project received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 722095.

For more information about the GlySign consortium and to access GlySign video please visit our R&D webpages.






Ludger Support - Resources webpages

Announcing the Launch of our New Support – Resources Webpages

Oxford – October 2020

We are pleased to announce the launch of the newly redesigned Support - Resources section of our website at www.ludger.com/support. The new webpages feature a modern design, improved functionality, and easy access to essential information such as application notes, presentations, product comparison tables, FAQs. Our goal with this new section is to provide our visitors an easier way to learn about our technologies, products, and services.

We will be constantly updating our content with helpful information. We hope you find the new webpages useful and easy to navigate.

For any questions, suggestions, feedback or comments, please contact us at info@ludger.com






Publication in Nature Communications: Prominent members of the human gut microbiota express endo-acting O-glycanases to initiate mucin breakdown

Oxford – September 2020

Ludger Nature Comm Publication - Figure 1A
Fig. A model of an intestinal mucin glycoprotein showing complexity and variability of O-glycan chains

The role of the human gut microbiota and its interactions with the gut wall, and more specifically the glycans covering proteins lining the gut wall, is a highly complex and under-explored one. Gaining a better understanding into the metabolic mechanisms of the microbiota will ideally lead to a better understanding, and potential therapeutic interventions of a number of diseases that affect people and their digestive system. This paper, led by Newcastle University came about through a number of collaborations of which Ludger played an instrumental part and satisfyingly connected our collaborators withing the Glycoenzymes for Bioindustries consortium, with those at Leiden University Medical College. The paper highlights how investigations of this nature can have both long term objectives, understanding disease mechanisms, and short term opportunities, namely discovery of glycanase enzymes that can be cloned and have industrial and other academic application.

Please visit our Procainamide webpage for more information on how to characterise glycans using LC-MS. And for more information about this article visit our Publications webpage.






Grant News: Innovate UK Business-led Innovation in Response to Global Disruption

Oxford – August 2020

Ludger GlyHeath-COVID - Fusion Medical Animation
Photo by Fusion Medical Animation on Unsplash

Ludger is pleased to announce being awarded an Innovate UK grant as part of the governments covid-19 response competition "Business-led innovation in response to global disruption round 2".

This project is to accelerate the development of a proof-of-concept prototype of GlyHealth-COVID - a medical technology we are designing to identify 'immunofrail' individuals who are at high risk of becoming severely ill or dying from COVID-19. Initially, it was thought that only the elderly would get very sick and die from COVID-19 but around 25% of deaths have been in those aged below 65 with severe sickness and death occurring even in young, apparently fit, individuals. Using current medical technology doctors cannot foresee which of their COVID-19 patients will have the worst health outcomes so they must guess. Our work is on early detection and prediction of the health trajectories of patients with chronic and acute inflammatory diseases (IDs). COVID-19 is an acute-ID which causes sickness and death in similar ways to chronic-IDs but at greatly accelerated speeds. Immunofrailty is a concept we have developed during over a decade of research. It is a very complex condition with a wide range of causes that all lead to high levels of chronic inflammation. Current medical technology can only measure acute inflammation, not immunofrailty-related chronic inflammation. We have developed a medical technology called GlyHealth that reliably measures immunofrailty and chronic-inflammation in individuals from changes in biomolecular patterns in their blood. This can be done without interference from acute inflammation. Those patterns are signals that can potentially predict dangerous inflammatory storms.

GlyHealth-COVID is based on a test that we have developed to help gastroenterologists caring for inflammatory bowel disease (IBD) patients. We can test its ability to identify immunofrail COVID-19 patients in an initial clinical study. If it performs then GlyHealth-COVID would be able to reliably identify vulnerable COVID-19 patients from small (0.2ml) samples of their blood. The samples can be taken with easy to-use fingerprick blood collection kits which can used at home.

For more information on our collaborative projects, please visit our R&D medical glycomics webpage.






New product: Sialidase Testing Panel

Oxford – Sept 2020

We are announcing the launch of a new product – Sialidase Testing Panel – a glycan standard containing a mixture of α2-3, α2-6 and α2-8 sialylated oligosaccharides:

  1. 3’-Sialyl Lewis X [Neu5Ac-α2-3Gal-β1-4(Fuc-α1-3)GlcNAc]
  2. Disialyllactose [Neu5Ac-α2-8NeuAc-α2-3Gal-β1-4Glc]
  3. Sialyllacto-N-tetraose a [Neu5Ac-α2-3Gal-β1-3GlcNAc-β1-3Gal-β1-4Glc]
  4. Sialyllacto-N-tetraose c [Neu5Ac-α2-6Gal-β1-4GlcNAc-β1-3Gal-β1-4Glc]
  5. Disialyllacto-N-tetraose [Neu5Ac-α2-3Gal-β1-3(Neu5Ac-α2-6)GlcNAc-β1-3Gal-β1-4Glc]
Ludger Sialidase Testing Panel Structures

Sialidase Testing Panel can be used as process positive control for sialidase digestions. Applied alongside the samples, it enables the analyst to test if the sialidase used has the required specificity and has worked correctly.

The Sialidase Testing Panel is available labelled with 2-AB or procainamide.
Cat# CAB-STP-NEUAC-01 / CPROC-STP-NEUAC-01

To find out how to incorporate the Sialidase Testing Panel into an exoglycosidase sequencing workflow and to view range of Ludger exoglycosidase enzymes available, visit our Exoglycosidase page.

For enquiries or more info, please contact: info@ludger.com






Publication in Scientific Reports: Insights into the salivary N‑glycome of Lutzomyia longipalpis, vector of visceral leishmaniasis

Oxford – August 2020

Successful collaboration between Ludger and The Liverpool School of Tropical Medicine, resulted in publishing an article in Scientific Reports titled “Insights into the salivary N‑glycome of Lutzomyia longipalpis, vector of visceral leishmaniasis." Ludger’s contribution included:

Ludger Scientific Reports publication - Figure 3B
Mass spectrometry analysis of released N-glycans from Lu. longipalpis salivary glycoproteins Positive-ion MS/MS fragmentation spectrum for m/z [727.8]2+.
Green circle, mannose; Blue square, N-acetylglucosamine; Proc procainamide
  • Help with the study design and strategy
  • Release of N- and O-glycans from sand fly saliva
  • O- and N-glycan profiling and characterisation using orthogonal techniques (UHPLC, LC-ESI-MS and exoglycosidase digestions)
  • Supporting the visual reporting of structures and nomenclature of glycans identified in the study

This is the first study of the salivary N-linked glycans of Lu. longipalpis, providing detailed information about their structures and relative abundances. Additionally, the discovery of a 144 Da (unknown) modification present in some salivary glycans is reported. This study provides new insights into how these structures could be recognised by vertebrate host cells.

This work was partially supported by funding from the Wellcome Trust and GlycoPar EU FP7 Marie Curie Initial Training Network.

To find out how to utilise enzymes in glycan characterisation visit our Exoglycosidase enzyme page. Visit our Procainamide webpage for more information on how to characterise glycans using LC-MS. And for more information about this article visit our Publications webpage.






Quantitative Glycan Standards: Strategies for building quality into your experiments using Ludger BioQuant (BQ) Standards

Oxford – August 2020

Robust analytical strategies are required to meet the challenge of accurately and reliably characterizing glycosylation. One way to meet this challenge is by integrating quantitative standards into your analytical processes.

Commercial quantitative glycan standards are key tools for:

  1. determining analytical process efficiency
  2. quantifying samples

The value of incorporating a quantitative standard into your workflow is that you know what the resulting data is expected to be – this allows you to assess the performance of your process.

If you have confidence in your process you can have confidence in your data.


Ludger’s Quantitative Glycan Standards BioQuant (BQ) Standards

Ludger’s range of quantitative standards are called BioQuant standards. BioQuant standards are designed to fit simply into your analysis workflow and you treat them as you would any other sample.

Ludger has integrated BQ quantitative standards into many of our analytical processes for glycan analysis and has set acceptance criteria. This allows the analyst to judge the results and know if they are truly in or out-of-specification.

BQ-GPEP-A2G2S2 as a Quantitative Process Standard (BQ-GPEP-A2G2S2-10U)

Ludger BQ-GPEP-A2G2S2 vial

The glycopeptide standard is comprised of an A2G2S2 glycan attached to the asparagine amino acid of a peptide with the sequence Lysine-Valine-Alanine-Asparagine-Lysine-Threonine (KVANKT).

BQ-GPEP is an integral process standard for assessing the performance of the quantitative assays. The two most common quantitative assays during glycan characterisation are monosaccharide and sialic acid analysis.

When using BQ-GPEP-A2G2S2 as a process standard in these quantitative workflows it is taken along with all other samples through the entire process; acid release and labelling. The amount of sialic acid is calculated using a calibration curve. Acceptance criteria have been set for the BQ-GPEP standard in both monosaccharide and sialic acid analyses, this allows the analyst to assess whether the release and labelling have worked as expected and with this information they can have confidence in their entire data set.

BQ-Chitotriose: Standard Used to Quantitate Other Analytes in HPLC/UPLC

Ludger BQ-Chitotriose vial

Chitotriose is a linear tri-N-acetylglucosamine glycan. It is available as an unlabelled glycan (BQ-CHITOTRIOSE-01) and is also available labelled with 2-AB or 2-AA dye (BQ-CAB-CHI-01; BQ-CAA-CHI-01).

BQ-Chitotriose quantitative standard is used as an internal standard, spiked into the sample to be quantified, run on an analytical platform for analysis and the quantity of glycan analyte is inferred by comparison of peak height or peak area to that of the standard. This is possible because the trisaccharide is smaller than most glycans and hence elutes before them.




BQ-Man8 Quantitative Glycan Standard (BQ-CN-MAN8-10U) – External Quantitative Standard in HPLC/UPLC

Ludger Man8 Quantitative Glycan Standard vial

The BioQuant Man 8 glycan is a purified and quantified glycan standard

BQ-Man8 quantitative standard is used as an external standard meaning that it can be used to quantify analytes within a sample set (can be used across multiple samples). It is run on an analytical platform for analysis and the quantity of glycan analyte is inferred by comparison of peak height or peak area to that of the standard.

One advantage of the BQ-Man-8 standard is that it is a typical glycan found in biopharma and biological samples and as such is a characteristic N-glycan sample (in size and chemical properties). It can also be used to assess process efficiency (for example: labelling efficiency).

Please visit our Glycan Standards page for a full listing of the Quantitative Glycan Standards we have available.

If you have any questions or to request a quotation, please contact: info@ludger.com






Announcing the Launch of a Search Function for Ludger Product Certificates

Oxford – July 2020

We are pleased to announce the launch of our brand-new webpage for Ludger Product Certificates Search with immediate effect.

A certificate of analysis (CofA) is a document indicating that the product meets its product specifications. The CofA includes test results for each individual batch of a product that is obtained using our quality control procedure.

We wanted to make it easier and faster to locate a CofA on our website. The new webpage gives you the ability to search for, view, and download CofAs. To retrieve a CofA, please enter the product batch (or lot) number found on the product label.

Ludger Product Certificate Search - Step by step

For a Certificate of Analysis/Conformity please visit: www.ludger.com/product-certificates

We sincerely hope you enjoy navigating through our new webpage and if you have any suggestions or feedback please E-mail us.






Ludger Glyco-Tools: System Suitability Standards

Oxford – July 2020

Ludger Glycan Standards and Controls - Summary Table

Ludger provides a range of system suitability standards for different applications including the analysis of sialic acids (SA), monosaccharides (Mono), N-glycans (N) and O-glycans (O). We offer purified unlabelled, 2-AA, 2-AB, APTS, procainamide labelled or permethylated glycan standards, quantitative monosaccharides and sialic acid standards, quantitative glycopeptide standards and glycoprotein standards.

They can be used for:

  • Process controls for release, labelling and analysis
  • System suitability testing for MS, (u)HPLC and CE
  • GU (glucose unit) calibration for (u)HPLC
  • Structure identification
  • Quantitation
  • Exoglycosidase sequencing (positive and negative controls)

Please visit our Glycan Standards page for a full listing of the system suitability standards and controls we have available.

If you have any questions or to request a quotation, please contact: info@ludger.com






Research & Development for Ludger Medical Glycomics programmes

Oxford – June 2020

Ludger Research and Development for Ludger Medical Glycomics

Dysfunctional glycosylation is associated with a wide range of diseases including cancer, diabetes and cardiovascular disease (CVD), as well as congenital, immunological and infectious disorders. This area of study originated in the work published in Nature by Parekh et al in 1985 entitled 'Association of Rheumatoid Arthritis and Primary Osteoarthritis with Changes in the Glycosylation Pattern of Total Serum IgG' (Nature 316, 452-457) and has led to a strong initiative to explore the relevance of glycomics in Precision Medicine.

Despite the strong evidence for measuring glycosylation in disease and biologicals, glycomics is significantly lagging behind genomics and proteomics, mainly due to the absence of high-throughput analytical methods which can reliably quantify a multitude of glycan structures in complex biological samples. Through internal investment and grant funding, Ludger has developed a high throughput glycan analysis platform, utilising kits and glycan standards developed and manufactured at Ludger, that enables the analysis of 96 well plates of glycoprotein samples. This has reliably allowed us to process and analysis thousands of serum and plasma glycomes to establish robust and repeatable methods to generate glycomics data. The glycomics data generated from our studies is being used to identify new biomarkers and for patient stratification in specific diseases.

Ludger's Medical Glycomics Programmes have been established to study changes in glycosylation in different body states. We are working in a number of well-established partnerships on a range of collaborative projects including working on IBD, cardiovascular disease, biomarkers for healthy ageing and, as part of the GlySign consortium, on glycan signatures for disease in MODY (Mature Onset Diabetes of the Young) and novel high throughout plate based methods.

If you have a precision medicine project that requires the use of glycosylation analysis to provide glycan biomarker data or would like to collaborate with us on a project that you have, please get in contact with us: info@ludger.com






Ludger Glyco-Tools: Quantitative Monosaccharide Analysis

Oxford – June 2020

Monosaccharide analysis is a regulatory requirement laid out in the ICH Q6B guidelines for characterisation of biopharmaceuticals. This information can be used at all stages of drug development as a method of determining the type of glycosylation (N-linked and/or O-linked) and the extent to which glycosylation has occurred. It can also be used to demonstrate consistency between batches for QC lot release during the manufacturing process.

Ludger Monosaccharide Analysis

A widely used method for monosaccharide analysis is as follows:

  • Release of monosaccharides from the glycoprotein by mild acid hydrolysis.
  • Fluorescent labelling of released monosaccharides with 2-aminobenzoic acid (2AA).
  • Relative quantitative analysis of 2AA-labelled monosaccharides by HPLC or UPLC.

Ludger offers a Monosaccharide Release and Labelling kit (Cat No. LT-MONO-96) for quantitation of GlcN, GalN, Gal, Man, Glc, Fuc, Xyl.

The kit contains reagents for up to 96 samples and a quantitative standard (CM-MONOMIX-10) containing 6 monosaccharides (GlcN, GalN, Gal, Man, Glc, Fuc) and Xyl standard (CM-XYL-100).

A key component of well-designed analytical strategy is inclusion of process standards.

Ludger offers several process standards for Monosaccharide analysis which will enable you to check the efficiency of glycan release, labeling and recovery and will give you confidence in the accuracy of your monosaccharide measurements:

We also offer two choices of column dependent on whether you are using HPLC or UPLC systems in your laboratory and HPLC/UPLC solvent for analysis of the labelled monosaccharides:

For information such as product specifications, companion products, storage, and stability visit our Monosaccharide Analysis page.

If you have any questions or to request a quotation, please contact us at info@ludger.com






Publication in Cellular and Molecular Life Sciences: Discovery of antennary fucosidase capable of removing fucose from sialylated glycans

Oxford – June 2020

Ludger - Publication in Mol Cell - Figure 4d
Figure 4D - Model of the orientation and conformation of sLeX bound to R. gnavus E1_10125 proposed by MD simulations

Successful collaboration between Ludger and The Gut Microbes and Health Institute Strategic Programme, Quadram Institute Bioscience resulted in publishing an article in Cellular and Molecular Life Sciences titled “Fucosidases from the human gut symbiont Ruminococcus gnavus".

As part of this research, we performed detailed characterization of the activity profile of the fucosidases isolated from gut bacteria R. gnavus, which was made possible by LC-MS analysis of procainamide labelled glycan products of enzymatic reactions. This study has contributed to our understanding of the adaptation of gut symbiont species to their ecological niches and has lead to a discovery of the novel fucosidase with unique specificity, capable of removing α1-3,4 fucose from sialylated glycan epitopes (like Sialyl Lewis X/A). The discovery will facilitate the future research into Lewis epitope-associated diseases like cancers and diabetes.

Characterised fucosidase will soon be available as part of Ludger exoglycosidase enzymes portfolio. Please enquire at info@ludger.com for more information.

To find out how to utilise the enzymes in glycan characterisation visit our Exoglycosidase enzyme page. Visit our Procainamide webpage for more information on how to characterise glycans using LC-MS. And for more information about this article visit our Publications webpage.






Ludger - Glycan Analysis Services webpages

Announcing the launch of the new webpages for Ludger Glycan Analysis Services

Oxford – May 2020

We are very excited at Ludger to launch our newly designed Glycan Analysis Services webpages with immediate effect.

The detailed knowledge of glycan structures is invaluable for a better understanding of their biological functions. Glycans can have a significant effect on the clinical safety and efficacy of biopharmaceuticals. Issues with glycans have caused great financial, legal and regulatory problems for those companies who have not dealt effectively with their product's glycosylation.

Ludger offers custom analytical services to suit your individual requirements and has been doing so since 1999. Ludger's Glycan Analysis Team has many years of expertise in analysing glycosylation from a variety of sample types including:

  • Biopharmaceuticals: monoclonal antibodies (mAbs), glycoprotein hormones (e.g. follicle stimulating hormone (FSH) and erythropoietin (EPO), Fc fusion proteins, vaccines
  • Cells: mammalian cell lines, bacterial cell components
  • Biological fluids, tissues and others

Ludger's Glycan Analysis Services include: Quantitative Sialic Acid Analysis, Quantitative Monosaccharide Analysis, N- and O-glycan profiling and characterization, site occupancy and site-specific glycosylation analysis.

The new webpages have been designed with an intuitive layout for ease of navigation and the menu options are designed to guide you to the most appropriate glycoprofiling programme.

We have incorporated valuable information regarding each glycoprofiling program/module.

For more information please visit the Glycan Analysis Services pages.
If you have any questions or to request a Study Proposal and quotation, please contact us at info@ludger.com or Dr Radoslaw Kozak directly at rad.kozak@ludger.com

We sincerely hope you enjoy navigating through our new Glycan Analysis Services webpages and if you have any suggestions or feedback please E-mail us.








Coronavirus disease 2019 (COVID-19)

Oxford – May 2020

The current global situation with COVID-19 has changed our daily lives and the way we interact. Ludger is driven by mutual objectives: staying safe and making critical updates to our laboratories, logistics, shipping, supply chain, purchasing and other processes to ensure you can purchase products for your glycoprofiling workflows or send Ludger samples for testing.

Some of our deliveries may take longer than usual and some of the products might not always be readily available. We hope you will bear with us as we do our best to continue to deliver to you during these challenging times.

As always, our Scientists are happy to help with technical support and can discuss suitable products for your needs as well as our custom Glycan Analysis Services.

We are here to support you.
Ludger






Coming Soon: Sialidase Testing Panel

Oxford – May 2020

Ludger - Sialidase Testing Panel
Figure: HILIC-UPLC profiles of procainamide labelled Sialidase Testing Panel (CPROC-STP-NEUAC-01). Top chromatogram shows undigested sample, bottom chromatogram shows sample treated with broad specificity sialidase (E-S001). Peaks were labelled with glycan structures. Arrows illustrate glycan digestion pathways.

A key component in a well-designed analytical strategy is the inclusion of standards and process controls. We are excited to announce the forthcoming launch of our Sialidase Testing Panel. This standard contains a mixture of α2-3, α2-6 and α2-8 sialylated oligosaccharides:

  1. 3'-Sialyl Lewis X
    [Neu5Ac-α2-3Gal-β1-4(Fuc-α1-3)GlcNAc] - contains α2-3 linked sialic acid along with branched α1-3 fucose (introducing steric hindrance for some enzymatic reactions)
  2. GD3 oligosaccharide - Disialyllactose
    [Neu5Ac-α2-8NeuAcα2-3Gal-β1-4Glc] - linear oligosaccharide containing α2-8 linked sialic acid
  3. LSTa; Sialyllacto-N-tetraose a
    [Neu5Ac-α2-3Gal-β1-3GlcNAcβ1-3Gal-β1-4Glc] - linear oligosaccharide containing α2-3 sialic acid attached to galactose
  4. LSTc; Sialyllacto-N-tetraose c
    [Neu5Ac-α2-6Gal-β1-4GlcNAcβ1-3Gal-β1-4Glc] - linear oligosaccharide containing α2-6 sialic acid attached to galactose
  5. DSLNT saccharide
    [Neu5Ac-α2-3Gal-β1-3(Neu5Ac-α2-6) GlcNAc-β1-3Gal-β1-4Glc] - branched sialic acid oligosaccharide

Sialidase Testing Panel can be used as process positive control for sialidase digestions. This standard enables you to test if the sialidase has required specificity and that it had worked correctly. Sialidase Testing Panel will be available 2-AB and procainamide labelled.
Cat# CAB-STP-NEUAC-01 and CPROC-STP-NEUAC-01

More information will be available soon on our website. To find out how to incorporate the Sialidase Testing Panel into exoglycosidase sequencing workflow and to view range of Ludger exoglycosidase enzymes available, visit our Exoglycosidase page.

For enquiries or more information, please contact: info@ludger.com






Publication in Frontiers in Chemistry: A MALDI-MS Assay for the Relative Quantitation of Antennary Fucosylated N-Glycans in Human Plasma

Oxford – April 2020

Ludger Publication - Frontiers in Chemistry

Ludger is delighted to announce the publication of a robust technique for analysing glycan biomarkers in patient samples developed by our PhD researcher Osmond Rebello whilst on secondment at Leiden University Medical College.

With the increase in health-related discoveries using glycan based biomarkers has come the need to utilise such biomarkers with an increase in efficiency and robustness using instrumentation that is rapid enough to analyse patient samples in timeframes suitable for meaningful clinical outcomes.

This paper addresses this need by demonstrating and validating a MALDI mass spectrometry technique for patient plasma glycome analysis coupled with linkage stabilisation and characterisation of sialic acids and analysis of antenarry fucosylation, frequently implicated in the progression of cancers.

The MALDI-MS instrumentation used was both high end type such as seen in specialist analytical laboratories, but also an instrument of more modest capabilities regularly found in routine analytical labs.

The paper describes in detail how to implement the technique and demonstrates its effectiveness against some colorectal cancer patient samples where antennary fucosylation levels were found to reduce after surgery, indicating success in removing cancerous tissue.

Please visit our Enzyme webpage for more information on how to release glycans from plasma samples, and for more information about this article visit our Publications webpage.






Enzymes for removal of intact glycans from glycoproteins and glycopeptides

Oxford – April 2020

Ludger Endoglycosidases - PNGase-F

Endoglycosidases are the most widely used group of enzymes for the analysis of glycoproteins. These enzymes release intact glycans from a wide variety of glycoproteins and are generally preferred to chemical glycan release methods due to their high efficiency and gentle mechanism of action.

Our featured products include:

  • PNGase F for the release of all types of N-linked glycans (high-mannose, hybrid and complex) from most glycoproteins and glycopeptides.

  • A range of other endo-acting enzymes for the release of N-linked glycans. This group of enzymes act by cleaving between the two innermost GlcNAc residues of the N-glycan core, leaving the terminal GlcNAc attached to the protein. These enzymes vary in specificities and include endoglycosidase F1, F2, F3 and H.

  • O-glycosidase for the enzymatic release of Core 1 O-linked glycans.

  • Ceramide glycanase for deglycosylation of glycosphingolipids.

For information such as product specifications, glycan cartoons depicting the activities of the enzymes, enzyme sources, enzyme specificities, storage and stability as well as images of the enzyme kits and kit contents visit our endoglycosidase page.

If you have any questions or would like to request a quotation, please contact us at info@ludger.com






New Product Launch: Ludger post exoglycosidase clean-up spin columns (LC-EXO-A6) and 96-well plates (LC-EXO-96)

Oxford – March 2020

Ludger LC-EXO-A6 and LC-EXO-96

We are excited to announce the launch of Ludger’s new post exoglycosidase clean-up spin columns (Cat # LC-EXO-A6) and 96-well plates (Cat # LC-EXO-96).

Exoglycosidases are used to support structural characterisation of glycans. Often, removal of enzymes from reaction mixture is needed before further processing and analysis of modified glycans.

At Ludger we offer a new product range that is designed for a rapid removal of enzymes and other protein material from glycan solution:

  • Post exoglycosidase clean-up spin columns – convenient spin tube format that is compatible with traditional laboratory centrifuge. Available in pack of 6. Cat # LC-EXO-A6

  • Post exoglycosidase clean-up 96-well plate – compatible with plate format centrifuge as well as vacuum manifold system (e.g. Ludger Cat # LC-VAC-MANIFOLD-KIT). Convenient if higher number of samples are required. Cat # LC-EXO-96

Both products: LC-EXO-A6 cartridges and LC-EXO-96 plates are suitable for unlabelled, 2-AB labelled, 2-AA labelled or procainamide labelled glycans.

To find out how to incorporate the clean-up consumables into exoglycosidase sequencing workflow and to view range of Ludger exoglycosidase enzymes available, visit our Exoglycosidase page.

For enquiries or more information, please contact: info@ludger.com






Grant news: Global Challenges Research Fund (GCRF) to trial innovations that aim to transform lives in developing countries

Oxford – March 2020

HPL GCRF grant

Ludger’s Human Performance Lab has been awarded a grant from Innovate UK for the GCRF to conduct a 6-months feasibility study on polycystic ovary syndrome (PCOS) in the Philippines. The purpose of the study is to assess the impact and commercial feasibility of an educational self-care programme for women with PCOS living in the Philippines. The project meets several of the UN Sustainable Development Goals including [1] Good health and well-being and [2] Quality education.

The study will run from April to September 2020, in collaboration with Masuna - a social enterprise and partner on the project based in the Philippines. The Ludger project team includes Dr Daryl Fernandes, Dr Christel Gudberg, and Gloria Mate.

For more information, visit: https://www.gov.uk/government/news/transforming-lives-in-developing-countries-apply-for-funding.






Final GlySign Consortium Event

Leiden, The Netherlands – February 2020

Ludger GlySign Consortium

The final GlySign consortium event, funded by the Marie Skłodowska Curie ITN EU grant programme, was hosted by Prof. Manfred Wuhrer at Leiden Medical Centre in Leiden, Netherlands.

The first two days of the week-long event were dedicated to an Industry and Academic Mobility workshop (InAMob) which was fully organised by the ESRs. They were able to share their experiences on time spent in both industry and academic environments with invited Masters and PhD students. Day one included presentations from three senior guest speakers, Dr Marija Mladic, Senior Associate Scientist Protein Analysis at DSM, Dr Anish Chakkumkal, Principal Scientist and Scientific Team lead at Janssen Vaccines, and Prof. Manfred Wuhrer, Professor Proteomics and Glycomics, LUMC. They discussed their career paths and shared valuable advice on transition and mobility between academia and industry.

The final symposium on day 3 focused on research into glycosylation signatures for precision medicine and included presentations from three guest speakers who are experts in the field, Dr Gestur Vidarsson (Sanquin Research and Landsteiner Laboratory, Amsterdam), Dr Mandy van Hoek (Erasmus MC, Rotterdam) and Dr Lucy Crouch (Newcastle University, UK). All the early stage researchers (ESRs) who are part of the consortium also shared their final results and further perspectives of their research. This included three of the ESRs that have been based at Ludger. Daniel Demus presented his work on the development of two techniques: an LC-MS/MS method and an enzymatic plate-based assay for the detection of HNF1A-MODY using a glycan biomarker. Osmond Rebello presented his work on the development of an enzymatic plate-based assay for the determination of galactose and sialic acid levels within glycosylated proteins and characterisation of novel enzymes for glycan structural determination. Alan Moran presented his work on the development of a novel chemical derivatisation method for separating sialylated N-glycans on C18 chromatography to determine specific sialic acid linkages. Dr Daniel Spencer (Head of Development) and Dr Richard Gardner (Lead Scientist), as supervisors from Ludger, also attended the meeting.

Day 4 focused on the annual updates of the project including updates on all work packages, deliverables, reporting and future work potentials.

The final day of the event was dedicated to a career development training session for the ESRs. This day included a number of career development modules lead by Craig Jackson (ESS Holdings), Helen Williamson (Horizons Unleashed) and Dr Daniel Spencer.

For more information about the GlySign consortium and our other collaborations please visit our R&D webpages.






Ludger Products in a Workflow

Oxford – February 2020

Ludger Products Flowchart

High quality analytical glycan profiling and characterisation has continued to present many challenges. Ludger, the leader in analytical technology for medical applications of glycobiology provides products for glycan monitoring and characterization.

Our Products Flowchart illustrates how Ludger products can be used in succession to release, clean up and derivatize glycans (and glycopeptides) from your glycoprotein samples ready for analysis.

Please visit our Products webpage to view this and other helpful presentations and resources.







A4B Network Meeting

Leiden, The Netherlands – January 2020

Ludger - A4B Network Meeting

Along with the scientific activities organized by the A4B project, Ludger was present at the last network meeting which was hosted by Dr Manfred Wuhrer at LUMC (Leiden, The Netherlands) on the 13th of January 2020.

The A4B consortium is funded by Horizon 2020 Marie Sklodoswka-Curie ITN EU project that aims for the development of improved analytical tools and purification methods for the qualitative and quantitative detection of therapeutic protein species and post-translational modifications.

The 15 early stage researchers (ESRs) taking part in this consortium, the principal investigators and members of the scientific board met to present and discuss the preliminary results of their research. Dr Richard Gardner (Lead Scientist) and Manuela Amez from Ludger attended this meeting and she presented her results towards the development of a novel and rapid labelling for N-glycan characterization and relative quantification.

This scientific meeting was followed by a 3-day training course organised by the A4B project on presenting scientific data for the ESRs.

For more information about the A4B consortium and our other collaborations please visit our R&D webpages.






NanoCarb autumn workshop

Royal College of Surgeons, Dublin, Ireland – January 2020

Ludger - NanoCarb August Workshop presentation

As beneficiary of the Nanocarb Project Ludger was present on the latest consortium meeting which was hosted by Dr. Marco Monopoli at the Royal College of Surgeons in Ireland from the 24th to 30th of November 2019. The Nanocarb funded by Marie Sklodowska-Curie ITN EU project that aims to produce high quality research of the potential health applications of glycosylated nanoparticles.

The 15 early stage researchers (ESR) taking part in this consortium attended to present preliminary results of their research. Javier M Melo from Ludger presented which is currently carrying out as part in collaboration with the development and glycan production teams at Ludger under the supervision of Dr. Daniel Spencer (Head of Development), Dr. Jenifer Hendel (Lead Scientist) and Simon Peel (Head of Glycan Production).

This event is part of the training schedule of NanoCarb, which aims to train a group of highly competitive researchers knowledgeable in both Carbohydrate chemistry and nanotechnology.

For more information about the NanoCarb project and our other collaborations please visit our R&D webpages.







CyberEssentials accreditation

Oxford – December 2019

Ludger - Cyber Essentials accreditation 2019

We are pleased to announce that for the second consecutive year since Oct 2018 Ludger has achieved Cyber Essentials accreditation. Cyber Essentials is a government-backed and industry-supported scheme to guide businesses in protecting themselves against cyber threats. This accreditation demonstrates Ludger’s commitment to security and our ability to defend both our organisation’s and customers’ critical data against prevalent cyber threats.








Carbomet Workshop – A roadmap for Glycoscience in Europe

Ludger - Carbomet - A roadmap for Glycoscience in Europe

Leiden, The Netherlands – December 2019

CarboMet held a workshop in Leiden jointly organised by Ludger (represented by Drs Daniel Spencer and Jenifer Hendel) and Leiden University Medical Centre to identify challenges and opportunities in glycoscience covering, biopharmaceuticals, diagnostics/precision medicine, microbiome and sustainable materials. The workshop was attended by 28 delegates from early career to more established researchers representing 10 EU member states.

The stakeholder consultation section of the workshop was led by Dr Daniel Spencer of Ludger Ltd. alongside Professor Sabine Flitsch of the Manchester Institute of Biotechnology and is being followed up by an online consultation to give the wider glycoscience community a chance to contribute to the next European roadmap ‘Glycoscience 4.0’ (you can view the current roadmap by clicking here). We encourage our clients to participate in this consultation.

Please visit the CarboMet homepage to access the survey: https://carbomet.eu







Ludger poster presentations from BioProduction Congress 2019 and Well Characterized Biologics 2019 now available online

Oxford – December 2019

Ludger Poster - Analysis of Glycosylation Critical Quality Attributes (GCQAs) of monoclonal antibody (mAb) therapeutics Ludger Poster - Targeted Analysis of Glycosylation Critical Quality Attributes from Glycoprotein Therapeutics

The following Ludger poster presentations are now available to view on the website:

  • "Targeted Analysis of Glycosylation Critical Quality Attributes from Glycoprotein Therapeutics"
    presented by Paulina Urbanowicz (Senior Scientist, Ludger) at BioProduction Congress 2019 in Frankfurt, Germany
  • "Analysis of Glycosylation Critical Quality Attributes (GCQAs) of monoclonal antibody (mAb) therapeutics"
    presented by Radoslaw Kozak (Head of Glycoprofiling and Interim Business Development Lead) at Well Characterised Biologics 2019 (WCB) in Reston (VA), USA.

The posters illustrate the case studies on how to identify and analyse key GCQAs during biopharmaceutical’s development and highlight the importance of implementation of process and system suitability standards in glycoanalytical workflows. The posters were well received and we would like to thank everyone who attended both events.

To view these and any of our other posters, please click on the miniatures or visit our Poster webpage.




Publication in Molecular & Cellular Proteomics: Interlab Study on Characterisation of N-glycans from NIST monoclonal antibody

Oxford – November 2019

Ludger Publication: Molecular & Cellular Proteomics

We are proud to announce the successful publication of a manuscript entitled 'NIST Interlaboratory Study on Glycosylation Analysis of Monoclonal Antibodies: Comparison of Results from Diverse Analytical Methods' in collaboration with National Institute of Standards and Technology, United States. Ludger’s contribution included:

  • Help with the study design and strategy
  • Analysis of the N-glycan profile of the reference standard antibody using orthogonal techniques (MALDI, UHPLC and LC-ESI-MS)
  • Supporting the visual reporting of structures and nomenclature of glycans identified in the study

The study provided a cutting edge view for glycosylation measurement of biologics and highlighted the need for harmonisation of analytical methods in the world of glycobiology.

Please visit the Ludger Products webpage for more information on our range of glycan characterisation techniques, and for more information about the article visit our Publications webpage.




Publication in Microbiology: Bacterial sialidase as a virulence factor in oral pathogen infections

Oxford – November 2019

Ludger Publication: Microbiology

A successful collaboration between Ludger and School of Clinical Dentistry, University of Sheffield resulted in publishing an article in Microbiology titled “Characterization of Porphyromonas gingivalis sialidase and disruption of its role in host-pathogen interactions”.

As part of this research, we performed detailed characterization of the activity profile of the sialidase isolated from oral bacteria P. gingivalis, which was made possible by LC-MS analysis of procainamide labelled substrates and products. This study has contributed to our understanding of the multifarious roles of bacterial sialidases in virulence by oral pathogens. Moreover, it indicated how sialidase inhibition with chemotherapeutics could be a promising strategy for periodontitis therapy.

Please visit our Procainamide webpage for more information on how to characterise glycans using LC-MS. And for more information about this article visit our Publications webpage.




Ludger at Well Characterized Biologics (WCB) 2019

Reston (VA) and New York Metropolitan Area, USA – November 2019

Ludger at Well Characterized Biologics (WCB) 2019

Dr Radoslaw Kozak (Head of Glycan Analysis Services) will be attending the Well Characterized Biologics meeting (WCB) on November 11-13th 2019 in Reston (VA), USA. Rad will be presenting a poster (“Analysis of Glycosylation Critical Quality Attributes (GCQAs) of monoclonal antibody (mAb) therapeutics”) and hosting a roundtable discussion titled “Glycosylation Critical Quality Attributes (GCQAs) for Biopharmaceuticals”.

Please contact us if you will also be attending and would like to meet during this conference. Rad will be also visiting our clients and partners in the New York Metropolitan Area, Wed & Thurs (13-14th November 2019) so if you would like him to stop by for a discussion that could be arranged.

If you would be interested in viewing this poster, please sign-up to our Glycotechnology News service to be notified when it becomes available.

For more information on the WCB 2019, visit: https://lifesciences.knect365.com/well-characterized-biologicals






Ludger at BioProduction Congress 2019

Frankfurt, Germany – November 2019

Ludger at BioProduction Congress 2019

Paulina Urbanowicz (Senior Scientist) will be attending the BioProduction Congress on November 4-7th 2019 in Frankfurt, Germany. She will be presenting a poster entitled "Targeted Analysis of Glycosylation Critical Quality Attributes from Glycoprotein Therapeutics".

Please contact us to let us know if you will also be attending and would like to meet.
If you would be interested in viewing this poster, please sign-up to our Glycotechnology News service to be notified when it becomes available.

For more information on the BioProduction Congress 2019, visit: https://lifesciences.knect365.com/bioproduction






Publication in PLOS One: Glycoanalytical technologies for cell line analysis

Oxford – October 2019

Ludger publication: PLOS One

A successful collaboration by groups at Ludger, Amsterdam UMC and Leiden University Medical Centre, as part of the GlyCoCan grant, resulted in publishing an article in PLOS One titled “Method comparison for N-glycan profiling: Towards the standardization of glycoanalytical technologies for cell line analysis”.

Studies into glycosylation changes in cancer have been performed on a variety of sample substrates. In contrast to the limited availability of tumour tissue and primary cells, the availability of in vitro models makes cell lines a suitable, and useful tool for studying biological mechanisms in disease.

With a number of techniques currently available for the analysis of protein N-glycosylation, this research provides an overview of the methodologies routinely employed for the release of N-glycans from in vitro established cell lines, highlighting the information that can be obtained from each and when they might be best used. Based on our most recent results indicating fast processing times, high sensitivity and repeatability, we demonstrate that the in-solution PNGaseF method, followed by procainamide labelling and UHPLC and ESI-MS, produces robust results, exhibits high throughput potential, and can be used as a standard approach for N-glycosylation analysis of in vitro established cell lines.

Please visit our Enzyme webpage for more information on how to release glycans from cell lines, and for more information about this article visit our Publications webpage.






How to optimize, measure and control Galα1-3Gal (non-human epitope) on your monoclonal antibody (mAb) therapeutics?

Oxford – October 2019

Ludger Application Note: Alpha-gal-containing biologics

The clinical safety and efficacy of a glycoprotein drug is significantly influenced by its glycosylation. For instance, the presence of terminal α1-3-linked galactose (alpha-gal) can affect the safety profile and lead to a potential adverse reactions and neutralisation of the drug, thus reducing its therapeutic efficacy. Consequently, regulatory authorities are have tightened the requirements for biopharmaceutical companies to characterise, control and monitor their therapeutics glycosylation. However, identification and quantitative analysis glycans can be difficult to achieve due to their complexity and heterogeneity.

Our new application note illustrates Ludger’s expertise and the range of glycan analysis services for detection of alpha-gal and how we can assist you with identifying the Glycosylation Critical Quality Attributes (GCQA’s) of your glycoprotein drug.

We offer alpha-Gal standards labelled with 2-AB [CAB-Alpha-Gal-01] and 2-AA [CAA-AlphaGal-01]. These standards can be used as positive controls in glycoprofiling sequencing experiments utilising alpha 1-3 galactose specific exoglycosidase.

To enquire regarding glycan analysis please contact rad.kozak@ludger.com or to place an order please contact info@ludger.com

For more information and to view the application note, please visit our webpage on glycan analysis.






Announcing the launch of the new webpages for Ludger enzymes

Oxford – September 2019

Ludger Exoglycosidase Enzyme Sequencing Example

We are very excited at Ludger to launch our newly designed Endoglycosidase and Exoglycosidase enzyme webpages with immediate effect. Glycosidases are essential enzymatic tools in glycan analysis workflows for glycan release and characterisation. Our featured products include:

  • PNGase F and range of other endo-acting enzymes for release of N-linked glycans from glycoproteins
  • O-glycanase for enzymatic release of O-linked glycans
  • Ceramide glycanase for deglycosylation of glycosphingolipids
  • Panel of exoglycosidases (including O-acetyl esterase, range of sialidases, galactosidases, fucosidase and mannosidases) that can be used in sequential digestions of glycans and glycoproteins and as tools for glycan structure elucidation

The new webpages have been designed with an intuitive layout for ease of navigation and the menu options are designed to guide you to the most appropriate enzyme of your choice for your glycan analysis workflows.

We have incorporated valuable information regarding each enzyme including product specifications, glycan cartoon depicting the enzyme activity, source of enzyme, enzyme specificity, storage, stability as well as images of enzyme kits and/or kit contents. Each enzyme page also has links to their associated product guides, certificates of analysis, specification sheets and safety data sheets along with companion products that are listed on the side bar to assist you with glycan profiling and characterisation.

For more information, regarding our new enzyme webpages please visit our Endoglycosidase and Exoglycosidase pages. If you have any questions or to request a quotation, please contact us at info@ludger.com.

We sincerely hope you enjoy navigating through our new enzyme webpages and if you have any suggestions or feedback please E-mail us.






Advantages of Ludger's Procainamide labelling technology versus rapid/instant tags

Oxford – September 2019

Ludger Advantages of Procainamide labelling technology versus rapid/instant tags Application Note

Choosing between labelling strategies for glycan analysis using liquid chromatography with fluorescence detection (LC-FLR) and mass spectrometry (MS) is a difficult task. Even though 2-aminobenzamide (2-AB) labelling has been a gold standard method for glycan profiling and characterisation using LC-FLR as an analytical platform, new contenders such as procainamide and rapid/instant tags have been in use recently due to their comparable fluorescence, increased MS sensitivity and short processing times (Keser T, et al. Front Chem. 2018;6:324).

However, it should be noted that procainamide labelling offers several advantages over rapid/instant tags. Our new application note outlines the features, benefits and advantages of Ludger's Procainamide labelling technology in comparison to gold standard labelling (2-AB) as well as other rapid/instant tags.

To view the application note and to learn more about Ludger’s Procainamide labelling technology please visit our webpage on procainamide labelling.

We offer procainamide labelling kits for labelling 24 samples (Catalogue numbers: LT-KPROC-24 and LT-KPROC-VP24) and 96 sample labelling kit (Catalogue number: LT-KPROC-96) formats. To enquire or place an order please contact info@ludger.com









Summary of HPLC and UHPLC columns for glycan analysis

Table of Ludger HPLC and UHPLC columns used for Glycan Analysis

Oxford – September 2019

At Ludger we offer a range of HPLC and UHPLC columns to suit your specific chromatographic applications.

The table summarises the different applications; column specifications including particle size, flow rates, column pressure, temperature and a link to the product guide to assist you with your chromatography analysis.

To view the pdf version of the table, please visit our Glycan HPLC and UHPLC columns and buffers webpage.

For any enquiries, technical advice or to request a quotation, please contact us at: info@ludger.com







LudgerSep Buffers for WAX and HPLC Analysis of Glycans

Oxford – September 2019

Ludger Buffers for WAX and HPLC Analysis of Glycans Table

LudgerSep buffers are produced to simplify the preparation of solvents for glycan analysis using either WAX columns or HPLC columns. The advantages of using our buffers include: ease of buffer preparation, consistence of pH for every batch purchased, there is no risk of contamination or out-of-specification problems that arise during glycan analysis when using LudgerSep buffers.

The table summarises the different applications; specifications including description, usage, storage and a link to the product guide to assist you with your glycosylation analysis workflow.

To view the pdf version of the table, please visit our Glycan HPLC and UHPLC columns and buffers webpage.

For any enquiries, to request a quote or place an order please contact: info@ludger.com







Ludger at Glyco25 2019

Ludger at Glyco25 2019

Milan, Italy – August 2019

Maximilianos Kotsias (Scientist, Ludger) will be attending the 25th International Symposium on Glycoconjugates 2019 August 25th-31st in Milan, Italy. He will be presenting a talk entitled “Advancements in glycoanalytical strategies for N- and O-glycan analysis from in vitro established cell lines”. Additionally, he will be taking part in a round table discussion regarding the use of mass spectrometry for the analysis of glycans.

Please let us know if you will also be attending the conference and contact us at info@ludger.com if you wish to meet with Maximilianos who can assist you with your N- and O-glycan analysis workflows.






Application note for uses of Glucose Homopolymer ladder in Glycan Analysis

Oxford – August 2019

Ludger Application Note - Glycose Homopolymer Standard

During glycan analysis using hydrophilic interaction liquid chromatography (HILIC) platform there are two main analytical goals: 1) to have a reliable and reproducible analytical system and 2) to identify your analytes with accuracy and confidence. Therefore, it is good practice to run a regular system suitability check and to implement reference standards within your analysis and at Ludger we use the Glucose Homopolymer (GHP) standard for our chromatographic glycan analysis.

Our new application note illustrates the uses of GHP standard as a system suitability standard and a reference standard to assist you with your chromatographic analysis.

We offer GHP standards labelled with 2-AB [CAB-GHP-30], 2-AA [CAA-GHP-30], and procainamide [CPROC-GHP-30]. To enquire or place an order please contact: info@ludger.com

For more information and to view the application note, please visit our webpage on glycan standards.






Ludger Procainamide Glycan Labelling - Presentations

Glycan labelling with Procainamide

Oxford – July 2019

We have developed a concise presentation that outlines the key features and benefits of Ludger's procainamide labelling technology. This is presented alongside a more detailed data presentation that focuses on specific N- and O-glycan workflows, case studies and examples of procainamide labelled human IgG, biosimilar Erythropoietin, and complex biological samples such as human saliva and Tsetse fly saliva.

Procainamide labelling permits glycan identification by either mass spectrometry or (U)HPLC, and because of its improved ionisation efficiency compared to 2AB labelling it can permit identification of minor glycans (‹1% relative peak area) by ESI-MS.

Ludger's procainamide labelling system is suitable for N-glycans, O-glycans, GSL-glycans, heparin or any sugar with a reducing terminus and uses the same reductive amination labelling method that has been used for 2AB & 2AA.

For more information on our procainamide technology and to view both these presentations, please visit: www.ludger.com/procainamide

To enquire pricing for procainamide labelling kits please email us at: info@ludger.com








CarboMet workshop

Leiden, Netherlands – July 2019

Ludger CarboMet 2019 workshop

Ludger organised a one day training workshop on ‘creating and using glycans in precision work’ in Leiden, NL on July 5th (following EuroCarb2019) as part of their involvement as an industry partner in the CarboMet consortium

The workshop was designed for young researchers and focused on the current state of commercial glycans used to support precision work. The classes, uses and manufacture of currently available glycan standards for qualitative and quantitative glycoanalysis work were discussed. Interactive exercises with the ‘value proposition canvas’ were used to explore how enhancing the repertoire and increasing access to commercially-available glycan standards would aid development and manufacturing in the biopharmaceutical and precision medicine bioindustry sectors. The workshop was led by Jenifer Hendel (Lead Scientist at Ludger Ltd) and Daniel Spencer (a member of the CarboMet organisational committee and Head of Development at Ludger Ltd).

For more information on the Glycan Standards we have available at Ludger, please visit our Glycan Standards page.






Ludger at Eurocarb 2019

Leiden, Netherlands – June-July 2019

Ludger poster - Strategic Implementation of Glycan Standards for Reliable Analysis of Glycomolecules - Eurocarb 2019

Dr Jenifer Hendel (poster ‘Strategic Implementation of Glycan Standards for Reliable Analysis of Glycomolecules’, presentation at Young Researcher (ECYR) Workshop and CarboMet workshop), Manuela Amez Martin (poster ‘A Novel and Rapid Strategy for Labelling N-Glycans and Glycopeptides for Qualitative Analysis Using UHPLC-ESI-MS’), Daniel Demus (poster, ‘Detection of HNF1A MODY Diabetes with Glycan Biomarker’), and Alan Moran (presentation, ‘Linkage-specific characterization of sialylated N-glycans from human plasma by LC-MS’) all recently participated at EuroCarbXX 2019 in Leiden, NL, June 30th to July 4th.

To view these and any of our other posters, please visit our Posters webpage.






Publication in Nature Microbiology: On breakdown of complex N-glycans by gut Bacteroides

Oxford, July 2019

Ludger Publication in Nature Microbiology - On breakdown of complex N-glycans by gut Bacteroides

A successful collaboration between Ludger and University of Newcastle through the Glycoenzymes for Bioindustries grant resulted in publishing an article in Nature Microbiology titled “Complex N-glycan breakdown by gut Bacteroides involves an extensive enzymatic apparatus encoded by multiple co-regulated genetic loci”.

This research has helped in understanding the breakdown of complex N-glycans by prominent Bacteroides species (human gut microbiota). The data revealed that the ability to utilize N-glycans is commonplace among gut Bacteroides due to an extensive enzymatic apparatus of specific endoglycosidases and exoglycosidases encoded in these bacteria. Detailed characterization of the N-glycan breakdown pathway, based on LC-MS analysis of procainamide labelled substrates provided significant insights into the mechanism of N-glycan degradation by key members of the microbiota and provided knowledge for future studies examining the role of this process in gut survival.

Please visit our Procainamide webpage for more information on how to characterise glycans using LC-MS.
And for more information on this article visit our Publications webpage.






Book Chapter Published In Synthetic Glycomes

Oxford, July 2019

Ludger Chapter in RSC Synthetic Glycomes - Current stage of commercially available glycans

Dr. Jenifer Hendel (Lead Scientist, Ludger Ltd) and Dr. Daryl Fernandes (CEO, Ludger Ltd) co-authored a book chapter in the book ‘Synthetic Glycomes’ published by the Royal Society of Chemistry. The book illustrates and provides a comprehensive review of the current state of the synthetic glycome and introduces the applications of the synthetic glycome. Chapter 17, written by Ludger authors is entitled ‘Current Stage of Commercially Available Glycans to Support Realisation of Biologic Drugs’.

This chapter emphasises the reasons why drug glycosylation is important, by providing case-studies of drug programmes which encountered serious complications that could have been avoided through better glycoanalysis approaches. Furthermore, the chapter summarises the different classes, uses and manufacture of currently available glycan standards for qualitative and quantitative glycoanalysis work. Finally, the chapter encapsulates how glycochemists could enhance the repertoire of commercially-available glycan standards in order to aid development and manufacturing of safer and more effective biologic drugs.

Please visit our Glycan standards webpage for more information on the standards we offer at Ludger and how you can use them.
And for more information on this chapter visit our Publications webpage.






Ludger at Young Researcher Symposium 2019

Madrid, June 2019

Ludger at VI Symposium of Medicinal Chemistry Young Researchers 2019

Manuela Amez Martin (Marie Skłodowska Curie Early Stage Researcher) will be attending the VI Symposium of Medicinal Chemistry Young Researchers June 21st in Madrid, Spain. She will be presenting a poster entitled “A novel and rapid strategy for labelling N-glycans and glycopeptides for qualitative analysis using UHPLC-ESI-MS”.

If you would be interested in viewing this poster, please sign-up to our Glycotechnology News service to be notified when it becomes available.

For more information on the Young Researchers Symposium, visit: https://www.seqt.org/en/events/young-research-symposium/.







Ludger DDW 2019 poster

Ludger at Digestive Diseases Week 2019

San Diego, May 2019

Archana Shubhakar (Business Development Lead and Senior Scientist, Ludger Ltd) presented a poster titled “Serum N-glycomic biomarkers predict treatment escalation in Inflammatory Bowel Disease (IBD)” at the Digestive Diseases Week 2019 in San Diego.

The poster highlights the prediction of future need for surgical intervention or escalation of medical treatment using glycomics biomarkers in IBD patients. The poster presentation was well received and we would like to thank everyone who attended the event.

To view this and any of our other posters, please visit our Poster webpage.

Watch this space to find out more about our exciting IBD biomarker discovery project or sign-up to our Glycotechnology News service for regular updates on this and other R&D and medical glycomics projects at Ludger.







NanoCarb

Milan, May 2019

Ludger DDW 2019 poster

The first annual NanoCarb consortium event funded by Marie Skłodowska Curie ITN EU project brought together expert principal investigators and early stage researchers (ESRs) to explore applications in advanced nanomedicine using Glyco-Nanoparticles. The event was hosted by Dr. Luigi Ley (University of Milan) and Laura Polito (The Institute of Molecular Science and Technologies).

The event was inaugurated with a session lead by Helen Williamson (Horizons Unleashed) by introducing the ESRs to a leadership framework and career development. A shared symposium with the ITN consortium GlycoVax (http://glycovax.eu/) over two days allowed speakers from both consortiums to use the opportunity to disseminate their work focusing on nanoparticle technologies and vaccine development. The NanoCarb ESR’s were given a platform to present to the consortium about their career journeys so far and the opportunity to summarize the focus of their upcoming projects for the next three years.

For more information on the NanoCarb project and our other collaborations, please visit our R&D webpages.






GlyCoCan

Ludger DDW 2019 poster

Amsterdam, May 2019

The final annual GlycoCan consortium symposium funded by Marie Skłodowska Curie ITN EU project was hosted by Prof. Dr. Govert Somsen and Dr. Ing. Sandra van Vliet at the Vrijie University in Amsterdam. The symposium focused on colorectal cancer research and included presentations from four guest speakers who are experts in the field. All the early stage researchers (ESRs) who are part of the consortium also shared their interesting research on colorectal cancer. Maximilianos Kotsias (ESR, Ludger Ltd) presented his work on the development of “Automated techniques for assessment of novel O-linked glycan-associated biomarkers from colorectal cancer tissues”. The event also included training for the ESRs which was managed by Dr Daniel Spencer (Head of Development, Ludger Ltd) and Helen Williamson (Horizons Unleashed), covering modules including stakeholder management (Helen Williamson) and a career development module lead by Craig Jackson (ESS Holdings).

We would like to thank everyone for their participation and help to make the event a great success.

For more information on our collaborative programs, please visit our Research and Development webpages.






Ludger participation at the 4th Canadian Glycomics Symposium

Banff, May 2019

Ludger sponsorship brochure at Canadian Glycomics Symposium

Ludger are very pleased to have helped with sponsorship for the 4th Canadian Glycomics Symposium May 15-19th in Banff, Canada, organised by the Canadian Glycomics Network (GlycoNet). As part of our participation we provided a brochure introducing the attendees to Ludger’s glycomics technology and services.






New Product Launch: Ludger Glycan Purification Cartridges

Oxford, May 2019

Ludger EC50 Clean up cartridges for glycan purification

We are excited to announce the launch of Ludger’s new glycan purification cartridges (catalogue no. LC-EC50-24). These cartridges have been designed to purify glycans from non-carbohydrate material including salts, proteins and detergents by electronic interaction of the glycans with the surface of the cartridge.

Purification of glycans can be performed using LC-EC50-24 cartridges in your workflow:

  • After enzymatic or chemical release of glycans from glycoproteins.
  • After exoglycosidases (enzymatic) digestion of glycans to release individual monosaccharides to confirm glycan identity and structure.
  • Before and after glycan labelling using fluorescent tags such as 2-aminobenzamide acid (2-AB) and 2-aminobenzoic acid (2-AA).

A wide range of glycans including N-linked and O-linked type oligosaccharides, tri-saccharides and larger structures can be purified using LC-EC50-24 cartridges and further analysed using orthogonal techniques.

For more information, please view our presentation by visiting: www.ludger.com/glycan-clean-up-ec50/

If you have any questions or to request a quotation, please contact us at info@ludger.com






CarboMet Workshop: Creating and Using Glycans in Precision Work

Leiden, July 2019

Ludger Carbomet Workshop

Ludger is organising a one day training workshop in Leiden, NL on July 5th (following EuroCarb2019) as part of their involvement as an industry partner in the CarboMet consortium (www.carbomet.eu)

The workshop will focus on the current state of commercial glycans used to support precision work. We will overview the classes, uses and manufacture of currently available glycan standards for qualitative and quantitative glycoanalysis work. Finally, we will summarise our thoughts on how glycochemists could enhance the repertoire of commercially-available glycan standards in order to aid development and manufacturing in the biopharmaceutical and precision medicine bioindustry sectors where the exploitation of carbohydrates will have huge impact. We will use interactive exercises which will include the introduction and use of the 'business model canvas'. The workshop will be led by Jenifer Hendel (Lead Scientist at Ludger Ltd) and Daniel Spencer (a member of the CarboMet organisational committee and Head of Development at Ludger Ltd).

For more information about the workshop and to register for this event, please visit: www.ludger.com/workshops/carbomet-july-2019/






A4B Leadership and Glycan Analysis Workshop

Oxford, March 2019

Ludger A4B workshop

Ludger Ltd and Horizons Unleashed Ltd. are very pleased to have run a greatly successful workshop for the Analytics for Biologics (A4B) Marie Skłodowska Curie consortium early stage researchers (ESR). The ESRs explored their natural leadership styles and planned for the future. They attended a series of lectures on the importance of the role of glycans in biopharmaceutical efficacy and safety and how to characterise them. In addition, they spent two days in Ludger’s training laboratory learning how to release glycans, fluorophore label and analyse samples using LC-MS systems. Feedback from the ESRs was excellent and we loved having them visit us!

The Analytics for Biologics consortium aims to train the next generation of independent thinking researchers in the science of producing the most effective and safe biopharmaceuticals. Ludger’s focus is on getting a deeper understanding of glycosylation profiles of biopharma drugs with an aim of producing faster and better methods of characterisation aiding the drug developers in shaping their biologics to improve both their safety and function.

For more information on the A4B project and our other collaborations, please visit our R&D webpages



Ludger A4B workshop logos











Top 10 Presentation Award to Ludger Scientist at ECCO Conference

Copenhagen, March 2019

© European Crohn’s and Colitis Organisation

Archana Shubhakar (Ludger Senior Scientist) attended the 14th European Crohn’s and Colitis Organization (ECCO) conference held at the Bella Centre in Copenhagen, Denmark.

Archana presented a talk entitled “Serum N-Glycomic Biomarkers Predict Treatment Escalation in Inflammatory Bowel Disease (IBD)” at the speed abstracting digital oral presentation. Her talk was well received and she won the best abstract presentation award for the ‘Mechanisms of intestinal inflammation’ session. The inset picture from the the 14th Congress of ECCO, shows Archana (second from right) receiving the award at the presentation ceremony.

Watch this space to find out more about our exciting IBD biomarker discovery project or sign-up to our Glycotechnology News service for regular updates on this and other R&D and medical glycomics projects at Ludger.






Ludger at RCSI Research Day and Irish Laboratory Awards

Ludger-Nanocarb Irish Laboratory Awards

Dublin, March 2019

Dr Daniel Spencer, Head of Development (Ludger) visited the laboratories of our collaborator Dr Marco Monopoli, STAR Research Lecturer at the Royal College of Surgeons in Ireland (RCSI) and attended the RCSI Research Day 2019. Marco presented an excellent talk entitled “Horizon 2020 European Training Network (ETN) ‘NanoCarb’“ where he described this new, exciting EU Marie Skłodowska Curie PhD programme. He also highlighted the collaborative work with Ludger focusing on glyco-production, chemoenzymatic modification of large molecules, nanoparticle handling and purification techniques.

In addition, Marco’s laboratory was nominated in the category of “Best Irish start-up laboratory” at the Irish Laboratory Awards 2019 - Gala event held at The Ballsbridge Hotel, Dublin. Marco and Daniel attended this gala event with the some of the best lab groups in Ireland. The inset picture shows Daniel (left) and Marco (right) networking and enjoying the gala event.

For more information on this and our other collaborations, please visit our R&D webpages







‘Implementation of Glycan Standards into a Glycan Analysis Workflow’ talk presented in Dublin, Ireland

Dublin, February 2019

ludger publications

Michael Butler the Chief Scientific Officer at the National Institute for Bioprocessing Research & Training (NIBRT) institute in Dublin invited Rad Kozak, Head of Glycoprofiling (Ludger) to present a talk on the importance and significance of glycan standards.

Rad’s talk focused on emphasizing the need to include and implement the use of system suitability standards, reference standards, process standards as well as quantitative glycan standards for robust and reliable glycosylation analysis. The talk was well received by the contract bioanalytical services group and the research group at NIBRT.

For more information regarding Ludger’s system suitability standards and controls please visit our
Glycan Standards webpages

And for any enquiries or to request a quotation, please contact us at: info@ludger.com






Publication: Improved and semi-automated reductive β-elimination workflow for higher throughput protein O-glycosylation analysis

ludger publications

Oxford, February 2019

A study by groups at Ludger and Leiden University Medical Centre as part of the GlyCoCan grant, has reported a largely automated system for high-throughput protein O-glycosylation analysis. Adapting reductive β-elimination release of O-glycans to a 96-well plate system allows the method to be performed on a liquid handling robot enabling characterization and relative quantitation of O-glycans from commercially available standards. The method, which was validated according to the ICH (Q2) R1 guidelines for the validation of analytical procedures, produced rapid and accurate data, and has the potential to be utilized for O-glycan characterization of biological samples, biopharmaceuticals as well as biomarker discovery.

This was recently published in PLOS One:
Kotsias M, Kozak RP, Gardner RA, Wuhrer M, Spencer DIR. Improved and semi-automated reductive β-elimination workflow for higher throughput protein O-glycosylation analysis. PLoS One. 2019;14(1):e0210759. Published 2019 Jan 17. doi:10.1371/journal.pone.0210759

For more information on this and our other publications, please visit our Publications webpage.






Publication: Colorectal cancer study using CRISPR technology

Oxford, January 2019

ludger publications

A study by groups at Ludger and Amsterdam UMC as part of the GlyCoCan grant, has demonstrated the altered expression of fucosyltransferases in colorectal cancer cells and its impact on N-glycosylation. Using CRISPR-dCas9-VPR technology to augment glycosyltransferase expression, resulted in a change of N-glycosylation at the cell surface. The findings show that exploitation of the CRISPR-dCas9-VPR system can provide a better insight into malignant cell transformation and how it is associated with tumor progression, metastasis and resistance to chemotherapy. This was recently published in Glycobiology.

Transcriptional activation of fucosyltransferase (FUT) genes using the CRISPR-dCas9-VPR technology reveals potent N-glycome alterations in colorectal cancer cells. Blanas A, Cornelissen LAM, Kotsias M, van der Horst JC, van de Vrugt HJ, Kalay H, Spencer DIR, Kozak RP, van Vliet SJ. Glycobiology. 2018 Nov 22. doi: 10.1093/glycob/cwy096

To view this and others, please visit our Ludger Publications webpage.










Updated clean-up and labeling technology tables

Oxford, January 2019

We have updated our LudgerClean and LudgerTag chooser tables to incorporate all of the clean-up and labeling technologies that we offer. This should make it easier to select the most appropriate kits for your needs but of course you can always contact us directly if you have questions.

ludger clean-up technology table ludger labeling technology table

Visit our Clean-up or Labeling featured pages to view the tables and more information






GlySign Grant mid-term review meeting and workshop

Oxford, December 2018

ludger glysign workshop

The Mid-Term EU review meeting and training workshop for the GlySign project took place at Leiden University Medical College (LUMC) from 10-14 December (Leiden, NL). This was attended by Early Stage Researchers (ESRs), Senior Researchers involved in the project from beneficiary institutes and partner organizations, the EU project monitoring officer and the EU appointed scientific reviewer. The ESRs gave oral presentations of their work to date during this session, provided feedback to the EU officer and appointed reviewer and finally the consortium received feedback from the same reviewers. In addition there were training workshops for the ESRs on the following topics:

Biologicals for the Clinic - (led by Ludger Ltd and LUMC) – the ESRs presented reviews of the commercialization of different classes of biologicals for clinical use, followed by round table discussions of each sector. Other areas covered included: how to investigate Erythropoietin glycosylation using HILIC-mass spectrometry of fluorophore labelled glycans, how human derived immunoglobulins are produced for use in the clinic at the Sanquin Institute and some of the essentials for designing a workflow for producing and analyzing a biological.

Quality Management (led by Ludger Ltd and Horizons Unleashed Ltd.) - They also learnt some of the aspects of assuring consistent and accurate effectiveness of biological assays with a review of the ICHQ2(R1) guidelines for validation of an analytical assay and learnt about efficient ways of root cause analysis to help them troubleshoot when things go wrong in the lab.

Scientific Writing (led by LUMC) – An informative session on writing a scientific paper and how to compose a scientific journal rebuttal letter.

For more information on our collaborative programs, please visit our Research and Development webpages.







PhD position available - NanoCarb

Oxford, December 2018

ludger nanocarb project

NanoCarb Project - Glyco-Nanoparticles for Applications in Advanced Nanomedicine - www.nanocarb.eu

Applications are invited for a PhD position on the Development of purification and modification strategies of N-type glycopeptide and glycan moieties for use as nanoparticle activity enhancers to be carried out under the supervision of Dr. Daniel Spencer at Ludger Ltd, Oxford, UK with the PhD registered with Dr Marco Monopoli at the Royal College of Surgeons in Ireland (RCSI). The project will focus on the purification of N-glycopeptides and N-glycans from natural sources in up to gram quantities. These compounds will then be modified using enzymatic techniques to incorporate 2-3 and 2-6 sialic acid linkages, including both NeuAc and NeuGc forms with O-acetylation. Additionally, modification of branched glycans to incorporate antennary fucosylation such as the sialyl-Lewis-X moiety will also be explored.

The research project will enable the candidate to greatly develop their experience in the fields of glyco-production, chemoenzymatic modification of large molecules, and purification techniques. In addition, a small portion of the project will allow the candidate to gain introductory experience in surface chemistry and nanoparticle handling. During the PhD programme, the candidate will carry out secondments with academic partners.

This is one of 15 PhD positions (Early Stage Researchers [ESRs]) open in the recently granted Marie Curie European Training Network (MC-ETN) project NanoCarb - Glyco-Nanoparticles for Applications in Advanced Nanomedicine. NanoCarb is a multidisciplinary consortium including universities, research centres and SMEs with broad expertise ranging from nanotechnology, carbohydrate chemistry, glycoprofiling and in vitro/in vivo screening specialists.

For more information on the project and how to submit an application, please visit our posting on findaphd.com:  https://www.findaphd.com/phds/project/marie-sklodowska-curie-esr-glycan-research-the-development-of-purification-and-modification-strategies-of-n-type-glycopeptide-and-glycan-moieties-for-use-as-nanoparticle-activity-enhancers/?p100793
Application deadline: Thurs 13th December, 2018

If you have any further questions please contact Daniel Spencer.






glyShape presentation - T1 cartridges

Oxford, November 2018

ludger t1 cleanup cartridges - glyshape presentation

We have produced an informative slide presentation, part of our glyShape series for biopharmaceutical glycosylation, to explain the features and benefits of using LudgerClean T1 cartridges in your workflow. LC-T1 cartridges can be used to clean up samples after labelling with 2-AB, 2-AA or APTS. Clean up is completed within 30min for a single sample; 2 hours for 96 samples. The presentation also includes a video with step by step instructions.

To view, please visit www.ludger.com/glyshape







Glycan Analysis Services

Ludger Glycan Analysis Services

Oxford, November 2018

Dr Louise Royle retired in October after being with Ludger for over 10 years.
We thank her for her hard work and dedication during this time and wish her well in the future. We are pleased to announce that Dr Rad Kozak will be taking her place as Head of Glycoprofiling. Rad says, “I am excited to take on this role and very much look forward to the challenge ahead. Our philosophy is to work with clients in a partnership offering dedicated tailored solutions to ensure a successful delivery of the required information. The services we provide can help answer questions about the structures of glycans at all stages of the drug lifecycle – from QbD studies to regulatory submissions, comparability studies,

Please contact us if you would like to find out more about our custom analytical services, and visit our Glycan Analysis webpage for more information.


Louise Royle, pictured with glycan analysis colleagues
Dr Jenifer Hendel and Dr Rad Kozak





CyberEssentials accreditation

Oxford, November 2018

Ludger Cyber Essentials Accreditation

We are pleased to announce that in October 2018, Ludger achieved Cyber Essentials accreditation. Cyber Essentials is a government-backed and industry-supported scheme to guide businesses in protecting themselves against cyber threats. This accreditation demonstrates Ludger’s commitment to security and our ability to defend both our organisation’s and customers’ critical data against prevalent cyber threats.







Prize winning poster at ICS 2018

Oxford, October 2018

ludger glycan standards poster

Dr. Jenifer Hendel attended the 29th International Carbohydrate Symposium in Lisbon, July 15-19th.

Jenifer’s poster entitled ‘Glycan Standards as Key Tools for Robust and Reliable Analysis of Glycoproteins’ was well received and she would like to thank everyone for the engaged conversations.

The journal of Organic & Biomolecular Chemistry also awarded it the Chemical Biology Poster Prize.

To view this and any of our other posters, please visit our Posters webpage













Introducing our glyShape presentation series

Oxford, September 2018

ludger glyshape presentation series

GlyShape presentations have been created for our clients in the biopharmaceutical industry to share Ludger's expertise and demonstrate how our systems can be integrated into labs as part of the drug development process. The series will provide useful overviews of glycan analysis methodologies and instruction guides for key technologies.

The first in this series introduces our V-Tag technology for N-glycan release and labeling. It includes a video that provides clear step-by-step instructions on how to perform the method using the LT-VTAG-C30 kit.

To view the presentation and for more information, please visit: www.ludger.com/glyshape

We'll have more presentations to follow. Sign-up to our Glycotechnology News service for regular updates.





New Product Launch -
V-Tag Glycan Release and Labeling Kit

Oxford, August 2018

ludger v-tag glycan release and labeling kit

We are excited to announce the launch of our V-Tag glycan kit which enables release of N-glycans from glycoproteins, N-glycan labeling and clean up within 1 hour for a single IgG type sample and 2 hours for 30 IgG type samples (timing for release by PNGase F might need to be adjusted for complex type samples). The glycans are then ready for analysis by (U)HPLC. Also, once labeled with V-Tag the glycans can be incubated with exoglycosidases to allow for structure assignment.

For more information and to view our presentation visit our
N-Glycan Labeling Feature Page

Cat # LT-VTAG-C30

If you have any questions or to request a quotation, please contact us at: info@ludger.com





Joint GlyCoCan and GlySign Business Process and Quality Management Workshop

Oxford, July 2018

ludger horizons unleashed glysign glycocan workshop ludger horizons unleashed glysign glycocan workshop

From 15-18th of May 2018 Ludger hosted the joint GlySign EID and GlyCoCan ITN workshop at the Culham Science Centre, UK. Course Leads: Daniel Spencer from Ludger Ltd and Helen Williamson from Horizons Unleashed Ltd.

Early Stage Researchers from the Marie Skłodowska Curie Actions projects GlySign EID and GlyCoCan ITN attended this workshop to learn about business processes such at the Business Model Canvas and the Value Proposition Canvas and explore how the tools can be useful in managing successful outcomes from their own PhD projects. In addition to this they also learnt about the value of robust quality management within an organisation. The workshop included seminars, interactive discussions, having a go at being a drinks manufacturer entrepreneur and a visit to a real food manufacturing company.

We would like to thank everyone for their participation and help to make the event a great success.

For more information on the GlySign and GlyCoCan projects, please visit our Research and Development for Medical Glycomics webpage.

ludger, horizons unleashed, glysign, glycocan





'New Publication in Glycoconjugate Journal on Automated glycomic profiling'

Oxford, July 2018

ludger glycoconjugate journal publication

A successful Business Interaction Voucher funded by IB Carb (www.ibcarb.com) was used to strengthen the glycomics collaboration between lmperial College London and Ludger Ltd. This has resulted in publishing the following article titled “Towards automation of glycomic profiling of complex biological materials” by Shubhakar et al. in the Glycoconjugate journal.

This paper compares the performance of an automated plate based method for N-glycan release and permethylation of glycans derived from mouse lung and kidney tissues to established standard glycomic protocols using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The study revealed that the automated workflow is highly repeatable and it shows highly comparable MALDI-TOF-MS N- and O-glycan profiles of complex heterogeneous glycoproteins when compared with the standard protocol.

The key advantages of using the automated sample processing steps include reduced sample preparation times, low sample volumes needed for processing, it eliminates the need for performing the labour intensive steps, minimises hands-on-time, increases sample throughput and this set up has also shown immense potential for multiplexing, making the automated samples processing method convenient, scalable, fast and reproducible.

Ludger’s LT-PERMET-96 kit was used for derivatising these complex biological samples. For more information on the LudgerTag Permethylation kits and to view the presentation on our technology, please visit our Permethylation webpage.

Pubmed link: Glycoconj J. 2018 Jun 16. doi: 10.1007/s10719-018-9825-8





'How well is your column performing?'

Oxford, June 2018

ludger application note on column performance

No matter what type of chromatography you are using (HILIC-LC for glycan GU, WAX-LC for glycan charge; RP-LC for sialic acid ormonosaccharide analysis) it is good practice to run a regular system suitability check.

Are you tracking your column performance?
If not, please see our Ludger Application Note for guidance.

And don't hesitate to contact us for more information at: info@ludger.com










Monosaccharide kit – new packaging

Oxford, May 2018

We are transitioning to new packaging for some of our kits, starting with our monosaccharide kit, LT-MONO-96. The new black boxes are shrink-wrapped and replace the plastic boxes (shown below, left).

We are excited about this new development as we believe this to be an improvement to the previous packaging. We have re-designed our labels and have also listed kit contents on the inside of the lid (with storage temperature guidance for each component).

Please contact us with any questions regarding.

Lugder Monosaccharide Kit - New Packaging Lugder Monosaccharide Kit - Interior Label






Glyshape seminars in Shanghai, China

Oxford, May 2018

Lugder Glyshape Seminar Shanghai, China Lugder Glyshape Seminar Shanghai, China

Daryl Fernandes, Chief Executive gave two seminars in Shanghai, China in March to representatives of biopharmaceutical companies and academia. The seminars were entitled “Ludger GlyShape Quality by Design”.

The seminars focused on how we are able to help companies develop biopharmaceuticals by understanding and optimizing glycosylation. Following his visit to Shanghai, Daryl visited clients in South Korea.

“It was a pleasure for me to visit Shanghai again and meet scientists from CFDA and biopharma companies across China. This was the second seminar we’ve held on the GlyShape China programme. It gave me an opportunity to further understand the needs of Chinese drug developers and regulators. I look forward to working closely with my colleagues over the coming years to help Chinese scientists develop and manufacture world-class biologic drugs that are safe, effective and affordable for patients and also very profitable for the companies.” - Daryl Fernandes

Please contact us if you wish to hear more about this.








Ludger at Euro Biosimilars Congress 2018

Oxford, April 2018

Lugder Procainamide Timeline

Archana Shubhakar (Senior Scientist) will be attending Euro Biosimilars Congress 2018 in Rome, Italy. She will be presenting a talk entitled, Automated permethylation for glycosylation analysis of biologics using MALDI-TOF-MS, and moderating Day 1 sessions.

Please let us know if you will also be attending and would like to meet.

For more information on Euro Biosimilars 2018 and the programme schedule, visit: https://biosimilars-biologics.pharmaceuticalconferences.com/europe/scientific-program





Procainamide Workflow - Sample preparation within 1 day

Oxford, April 2018

Lugder Procainamide Timeline

We have added the following information to our procainamide webpage and presentation:

  • An updated workflow for sample preparation (N-or O-glycan analysis)
  • Timings for N-glycan analysis of 96 samples
  • 4 peer-reviewed papers using this technology
  • A full list of products to use when performing N- or O-glycan analysis

To view the updated webpage and presentation please visit: www.ludger.com/procainamide





Analysis of Intact Released Glycans

Lugder Glycan AnalysisOxford, March 2018

We have recently added a short slideshow on intact released glycan analysis to our website. This demonstrates our analytical approach, with example workflows and data for IgG and EPO.

Please take a look by visiting our Glycan Analysis webpages











ISO accreditation

Lugder ISO 9001:2015Oxford, March 2018

We are pleased to report that Ludger has achieved accreditation to ISO9001:2015. This is the latest standard specifying requirements for a quality management system when an organization:

a) needs to demonstrate its ability to consistently provide products and services that meet customer requirements, and
b) aims to enhance customer satisfaction through the effective application of the system, including processes for improvement of the system and the assurance of conformity to customer requirements.

For more information, visit our 'About Us' page





‘IgE glycoforms’ paper published in Journal of Allergy and Clinical Immunology

Oxford, March 2018

Ludger IgE JACI publicationFrom our collaboration with Kings College London, as part of the ‘GlycoShape: Defining and designing altered IgE glycoforms’ project, we are pleased to announce the forthcoming publication of a paper in the Journal of Allergy and Clinical Immunology.

This paper describes the development of a tool to produce therapeutic IgE at high yields. Anti-tumour IgE has been demonstrated in a number of studies but it has been a challenge to produce of high enough yields for preclinical and clinical studies. Once the high yield recombinant IgE was produced, analyses at Ludger were performed to assess the glycosylation profile, and these revealed the importance of Man5 glycans in antibody binding and functionality. This study, using IgE as an example, has demonstrated how novel antibody therapies can be developed with enhanced effector functions.

Citation: Crescioli S, Chiaruttini G, Mele S, Ilieva KM, Pellizzari G, Spencer DIR, Gardner RA, Lacy KE, Spicer JF, Tutt ANJ, Wagner GK, Karagiannis SN. Engineering and stable production of recombinant IgE for cancer immunotherapy and AllergoOncology. J Allergy Clin Immunol. 2018 Jan 19. pii: S0091-6749(18)30081-2. doi: 10.1016/j.jaci.2017.12.986. [Epub ahead of print]

Article link: www.jacionline.org/article/S0091-6749(18)30081-2/fulltext





PhD position - Automatable and high throughput techniques to determine site specificity and quantification of N and O glycan profiles of therapeutic proteins

Oxford, February 2018

Ludger logoThe graduate school Analytics for Biologics (A4B) is offering 15 PhD positions with the aim to produce the much-needed specialists in field of production, down-stream processing and analysis of therapeutic proteins (TP). This Marie Sklodowska-Curie Innovative Training Network (ITN) is composed of 11 European academic and industrial institutions and provides the optimal scientific network for training schools and individual research projects toward a common goal.

One of the A4B PhD positions will be based at Ludger Ltd, funded and based at Ludger Ltd. for three years and supervised by Dr Daniel Spencer, with the PhD registered at the Leiden University Medical Centre with Professor Manfred Wuhrer as a four year course. The PhD position will suit someone with both synthetic chemistry and analytical chemistry experience and will involve the synthesis of a novel fluorophore to be used for automated high throughput glycan and glycopeptide analysis.

For more information and to submit an application, please visit: https://www.findaphd.com/search/ProjectDetails.aspx?PJID=90528





Ludger at WCBP 2018

Oxford, February 2018

Ludger Overview of Glycan LabelingDr. Rad Kozak and Dr. Claire Morgan have returned from successful participation at the 2018 CASSS WCBP Symposium in Washington DC.

Their poster presentations on Biopharmaceutical FSH characterisation and Sialylation in Biotherapeutics were well received and they thank everyone for the engaged conversations. They will shortly be sending each of the contacts made at the meeting a copy of the poster.

You can view their posters by visiting our Biopharmaceutical R&D webpage.





Choosing a Glycan LabelLudger Overview of Glycan Labeling

Oxford, January 2018

Our flowchart outlines the choices you can make when selecting a glycan label for your analytical detection method.

To view, visit our Glycan Labelling page which gives an overview of all of the Ludger technology available to you and their applications





Velocity System for Sample Clean Up

Oxford, January 2018

Ludger Velocity System for Sample Clean UpUse of a vacuum manifold for sample clean up speeds up processing times. Ludger’s Velocity vacuum manifold system is compatible with cartridges or plates, and is a valuable tool for your lab.

If you currently use our LC-S cartridges to clean up labelled glycans, you could try our LC-T1 cartridges with the manifold system and process 96 samples in under an hour. We have two plate systems which are compatible with the manifold system; our LC-PBM-96 plate can be used to clean up samples after treatment with endoglycosidase (PNGaseF) or exoglycosidases, and our LC-PROC-96 is designed for glycan clean up after procainamide labelling. The LC-permet-96 clean-up plate is used to enrich N-glycans prior to performing permethylation of released glycans.

We have summarised the options available in a table, which you can view on our Velocity Product page





Using Procainamide for Glycan Labeling

Oxford, December 2017

Ludger Procainamide Comparison TableWe have developed a table that outlines the features and benefits of Ludger's procainamide labeling technology in comparison to competitor offerings.

Procainamide labeling permits glycan identification by either mass spectrometry or (U)HPLC, and because of its improved ionisation efficiency compared to 2AB labeling it can permit identification of minor glycans (<1% relative peak area) by ESI-MS.

Ludger's procainamide labeling system is suitable for N-glycans, O-glycans, GSL-glycans, heparin or any sugar with a reducing terminus and uses the same reductive amination labeling method that has been used for 2AB & 2AA.

For more information, please visit our Procainamide webpage.





Application Note for Quantitative Sialic Acid Analysis

Oxford, November 2017

Ludger Application Note - Sialic Acid AnalysisSialic acid analysis is a regulatory requirement laid out in the ICH Q6B guidelines for characterisation of biopharmaceuticals. We have produced an Application Note detailing how to perform quantitative sialic acid analysis using our technology. This includes the methods for sample preparation using the LudgerTag DMB release and labelling kit and our system suitability standards as well as information for LC analysis using LudgerSep (U)HPLC columns.

To view, please visit our webpage on Sialic Acid Analysis











Ludger appoints Paul Plume as Chief Operating Officer

Oxford, September 2017

Ludger logoLudger Ltd, providing world-leading glycobiology expertise, glycotechnology products and services, is delighted to announce the appointment of Paul Plume as Chief Operating Officer

Paul joins Ludger in early November. His previous role was Program Director at the Getinge Group where he worked on stakeholder Group Projects spanning 6 global manufacturing sites, 3 global design centres and all group functions. Projects covered operational, engineering, regulatory, computer system, quality, branding and product safety. Prior to this Paul was at Abbott for over 12 years moving through a variety of functional areas covering capacity planning, engineering and operations for a variety of technical, operational and sales units within Abbott's Diabetes Care division. He has a Bachelor in Engineering from Leeds and studied on the Advanced course in Design, Manufacture and Management (ACDMM) from University of Cambridge.

Paul will add great value to Ludger as it continues on its journey.





China BioPharma Seminar

Oxford, September 2017

Daryl Fernandes presenting in ShanghaiDaryl Fernandes, Chief Executive of Ludger Ltd gave a seminar to members of the Chinese Biopharmaceutical Industry on September 15th 2017. This event was an excellent opportunity for us to meet drug developers in China and to share our expertise in the field of Glycobiology. The event was held at Shanghai‘s Zhangjiang Hi-tech Park Conference Hall. Daryl’s talk was entitled "GlyShape: a strategic technology programme for streamlining QbD-based development of glycoprotein therapeutics" and attracted much interest.

Lily Wang, Technical Sales Executive, Ludger China said "This seminar was a great success and there were many interesting technical discussions on the day". Cindy Li, Sales Executive, Ludger China commented "We look forward to working closely with our clients in China and helping them to achieve their goals".

Bruker Daltonics co-hosted this event and gave a talk entitled "Bruker Innovative Mass Spectrometry helps you improve the efficiency of protein drug mass spectrometry".

For more information on Ludger's GlyShape programme and how it can be implemented for biosimilar drug development please contact us: info@ludger.com





PNGaseF enzyme for more rapid glycan release

Oxford, September 2017

Ludger-NEB partnership

In partnership with New England Biolabs®, (NEB®), we are selling a PNGaseF enzyme kit which is suitable for releasing glycans from as many as 150 samples. The kit (Cat No. LZ-rPNGaseF-kit) contains 75,000 units of recombinant PNGase F at concentration of 500,000 units/ml.

PNGase F is suitable for release of all types (high-mannose, hybrid and complex) N glycans from glycoproteins and glycopeptides under denaturing or non-denaturing conditions. Supplied glycerol free for optimal performance in HPLC and MS intensive methods.

Cat #    LZ-rPNGaseF-kit

For enquiries or more information, please contact: info@ludger.com





Horizon 2020 Grant to Study EPO and TNF-AB

Oxford, September 2017

Ludger-Marie CurieLudger is a member of a pan European consortium which has been awarded H2020-MSCAITN-2017 (Marie Skłodowska-Curie Innovative Training Networks) Grant focussing on the qualitative and quantitative analysis and purification of therapeutic proteins. This 4-year project begins in October and a PhD student (ESR) will be based at Ludger for three years.

Our PhD student will focus on the development of automatable and high throughput techniques to determine site specificity and quantification of N and O glycan profiles of EPO and TNF-AB. The project will build upon a recently developed product, VTAG, used to analyze and relatively quantify glycan types on the Fc receptor of monoclonal antibodies without releasing the glycan.

Glycans from the therapeutic proteins will be analysed using UHPLC based hydrophilic interaction chromatography, and a glycopeptide fluorophore label will be optimized for MS and CGE-LIF analysis by chemical modification. Additionally, quantitative glycopeptide standards will be prepared and finally the assay will be automated and validated according to ICHQ2(R1) guidelines.





Characterisation of glycosylation in EPO using acetyl esterase

Oxford, July 2017

A paper entitled "Analysis of three epoetin alpha products by LC and LC-MS indicates differences in glycosylation critical quality attributes, including sialic acid content" has been accepted for publication by Analytical Chemistry. This work was the result of a collaboration between the University of Reading, University of Sheffield and Ludger.

Author(s): Thomson, Rebecca; Gardner, Richard; Strohfeldt, Katja; Fernandes, Daryl ; Stafford, Graham; Spencer, Daniel; Osborn, Helen. Reference: Anal Chem. 2017 Jun 9. doi: 10.1021/acs.analchem.7b00353. [Epub ahead of print] PMID: 28509534

LudgerZyme Acetyl EsteraseAs part of this work, we used acetyl esterase (sialate-O-acetylesterase) to remove 9-, 8- and 7-O-acetyl groups from the EPO biopharmaceutical glycans as these sugars and their acetylation are believed to be essential factors for the function, efficacy and half-life of the drug in patients. This enzyme can also be used for the characterisation of other highly sialylated biotherapeutics such as FSH and blood clotting factors.

Reference: Biochem J. 2015 Dec 1;472(2):157-67. doi: 10.1042/BJ20150388. Epub 2015 Sep 16.

Acetyl esterase (sialate-O-acetylesterase) is available to order from Ludger:

Cat #

LZ-ACASE-KIT     Kit containing enzyme and buffer sufficient for 50 samples





Tools for Analysis of Negatively Charged Glycans

Oxford, May 2017

LudgerSep C bufferNegatively charged glycans (sialic acids, sulphated or phosphorylated sugars) often play a critical role in the function of a glycoprotein. For example sialic acids increase the serum half-life of glycoproteins by protecting them from degradation by the asialoglycoprotein receptor; sulphated glycans are involved in cell adhesion; and Mannose-6-Phosphate is a key targeting signal for transport of glycoproteins to lysosomes and is present in therapeutic enzymes (enzyme replacement therapies) developed for treatment of lysosomal storage diseases.

The LudgerSep-C3 column (Cat # LS-C3-7.5x75) is a weak anionic exchange (WAX) HPLC column that enables you analyse negatively charged sialylated, phosphorylated and sulphated glycans. This technique is also known as ‘charge profiling’. An example of the information that can be provided is the relative amounts of sialylation (1, 2, 3 or 4 sialic acids) on your glycoprotein which is important to know when analysing a highly sialylated protein such as erythropoietin (EPO).

Although sialylated and sulphated glycans can be separated by anion exchange at a low pH of 4.4, the phosphorylated sugars would not be fully charged and there would be multiple species in solution. In order to have one buffer which is suitable for separation of all anionic glycans (sialic acids, phosphorylated and sulphated sugars) Ludger recommends the use of pH9 ammonium formate buffer. The LudgerSep C buffer is a pH 9 ammonium formate buffer concentrate (Cat# LS-C-BUFFX4) which is suitable for separation of all anionic glycans (sialic acids, phosphorylated and sulphated sugars). This concentrate can easily be diluted with water and acetonitrile then used directly as a solvent for the LudgerSep C3 column.

Cat #

LS-C3-7.5x75     LudgerSep-C3 column
LS-C-BUFFX4      LudgerSep C buffer concentrate

For enquiries or more information, please contact info@ludger.com





Analysis of Erythropoietin (EPO)

Oxford, March 2017

Ludger EPO AnalysisErythropoietin (EPO) biologics generate substantial revenue for the biopharma industry and as their patents expire the market for EPO biosimilars is set to grow considerably. Analysis of the glycosylation of EPO is a challenging requirement for drug developers because this glycoprotein hormone exhibits significant heterogeneity and its high degree of sialylation and accompanying acetylation can significantly affect its therapeutic properties (particularly the circulation half-life).

At Ludger we have developed accurate and reliable methods for analysis of EPO. In the poster attached here (WCBP 2017) we focus on sialic acid analysis and demonstrate the use of our DMB sialic acid technology (LudgerTag DMB kit, LT-KDMB-A1) to obtain information on the relative levels of the N-acetyl, N-glycolyl and O-acetyl sialic acids. This information can be used in QC to monitor batch-to-batch variation, or for comparability studies.

We can also execute detailed characterisation studies of EPO using LC and MS to give you the information you require.
This includes site specific glycosylation analysis to determine the following:

  • percentage site occupancy for each N-glycan and O-glycan site
  • glycan profile for each glycosylation site, with GU and relative proportions
  • identification of glycans at each glycosylation site (by comparison to structures identified from released glycan analysis)

If you would like more information on the DMB sialic acid technology or our glycoprofiling services (including method transfer) please contact us: info@ludger.com





Senior Scientist awarded PhD

Oxford, January 2017

Ludger Leiden Kozak PhD defenseWe are delighted to announce that our Senior Scientist Radoslaw Kozak has successfully defended his PhD thesis at the Leiden University Medical Center, The Netherlands.

Rad has been with Ludger since 2008 and for the past 6 years Ludger has supported his PhD which was undertaken in collaboration with the University of Leiden. His thesis is entitled: 'Rapid and sensitive methods for the analysis and identification of O-glycans from glycoproteins'. This work has led to improvements in Ludger's methods and technology for O-glycan analysis.

We all congratulate our new 'Dr' on his achievement.

Full thesis available online: https://openaccess.leidenuniv.nl/handle/1887/45434

Rad (seated centre, with paranymphs Stephanie Holst and Albert Bondt)
Photo by Gerhild Zauner





Innovate UK Grant: MODY diabetes

Oxford, January 2017

Innovate UK LogoLudger is delighted to have been awarded the Biomedical Catalyst 2016 - Feasibility Study grant funding from Innovate UK, for a glycomics precision diagnostic assay for Maturity Onset Diabetes of the Young (MODY).

MODY affects 1-4% of the diabetes patient population and it is estimated that at least 90% MODY patients are misdiagnosed and therefore, often prescribed ineffective treatment. A clinical diagnostics ‘MODY’ assay has been developed to identify patients with the most common form of MODY, HNF1A-type. These patients have reduced plasma outer arm fucosylation which is caused by defects in the HNF1A gene. The MODY assay is a plate based biochemical assay which provides a faster and more affordable alternative to genetic testing.

This exciting project, named GlycanDx-MODY, will be led by Ludger Ltd and includes the following partners; Genos (Croatia) and OCDEM (Oxford University, UK). The goal is to develop a clear business and technology plan for this assay to facilitate its incorporation into a diagnostic pathway for MODY which will ultimately be adopted by healthcare providers at the primary care level.





New publication in Nature Communications on Inflammatory Bowel Disease

Oxford, January 2017

Ludger IBD Nature CommunicationsAn article entitled ‘Integrative epigenome-wide analysis demonstrates that DNA methylation may mediate genetic risk in inflammatory bowel disease’ has been published in Nature Communications. Ludger is a member of the IBD-Biom consortium which contributed to the work.

Citation: Integrative epigenome-wide analysis demonstrates that DNA methylation may mediate genetic risk in inflammatory bowel disease. N. T. Ventham, N. A. Kennedy, A. T. Adams, R. Kalla, S. Heath, K. R. O'Leary, H. Drummond, IBD BIOM consortium, IBD CHARACTER consortium, D. C. Wilson, I. G. Gut, E. R. Nimmo & J. Satsangi. Nature Communications 7, Article number: 13507 (2016); doi:10.1038/ncomms13507.

Article link: http://www.nature.com/articles/ncomms13507












Grant news: GlySign

Oxford, November 2016

Ludger - GlySignWe are delighted to announce that the GlySign project has begun. GlySign is a research training network on glycomic clinical markers and assay development for Precision Medicine. This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 722095

We are joined in the GlySign consortium by Genos and Leiden University Medical Center, and will shortly be recruiting two PhD students to be based at Ludger.

For more information please visit: http://www.glysign.eu/

If you are interested in applying for a PhD position please visit: http://www.glysign.eu/phd-projects/





Release of O-linked glycans from glycoprotein therapeutics

Oxford, October 2016

Ludger Orela KitLudger offers two Ludger Liberate kits that can be used to release of O-glycans from glycoprotein therapeutics; the Hydrazinolysis kit, LL-HYDRAZ-A2 and the Orela kit, LL-Orela-A2. Whilst hydrazinolysis is the gold standard method to remove all O-links, the Orela Kit contains reagents which are safer and much easier to handle.

O-glycans released using either of these kits have free reducing termini so are compatible with reducing-end labeling using reagents such as 2-aminobenzamide (2AB), 2-aminobenzoic acid (2AA) and Procainamide (Proc) allowing high-performance liquid chromatography (UHPLC) with fluorescent detection.

The Orela kit (Cat # LL-Orela-A2) can be used for up to 12 samples. Each kit includes LC-CEX cation exchange cartridges for O-glycan purification prior to fluorescent labeling.

The hydrazinolysis kit (Cat# LL-HYDRAZ-A2) can be used for up to 12 samples. The release conditions can be optimized for release of N-glycans, O-glycans or both N- and O-glycans. Each kit includes LC-CEX cation exchange cartridges for O-glycan purification and also LC-EB20 cartridges for N-glycan purification.

Ludger Liberate glycan release kits can be incorporated into a workflow for O glycan analysis in your labs:


n.b. exoglycosidase sequencing can be used to gain more detailed information on the O-glycan structures





New publication in Analytical Chemistry on High Throughput Permethylation

Oxford, August 2016

Ludger Permethylation Analytical ChemistryLudger has published an article in Analytical Chemistry presenting an automated and high throughput (HT) glycan sample preparation and permethylation method used for characterisation and relative quantitation of glycans, glycoprotein standards and biopharmaceutical samples (IgG4 and rhEPO). To our knowledge, this is the first largely automated workflow for permethylation that has been executed using a liquid handling robot and a commercially available kit. The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) profiles obtained showed good agreement with hydrophilic liquid interaction chromatography (HILIC) and ultrahigh performance liquid chromatography (UHPLC) data (which is referred to as the gold standard method for glycan analysis). Automated permethylation and extraction of 96 glycan samples was achieved in less than 5 h and automated data acquisition on MALDI-TOF-MS took on average less than 1 min per sample, making this method convenient, fast, and reliable.

Citation: Automated High-Throughput Permethylation for Glycosylation Analysis of Biologics Using MALDI-TOF-MS. Archana Shubhakar, Radoslaw P. Kozak, Karli R. Reiding, Louise Royle, Daniel I. R. Spencer, Daryl L. Fernandes, and Manfred Wuhrer. Analytical Chemistry Article ASAP; DOI: 10.1021/acs.analchem.6b01639.

Pubmed link: http://www.ncbi.nlm.nih.gov/pubmed/27479043





Hydrophilic interaction amide clean up cartridges

Oxford, July 2016

Ludger Scientific ReportsLudger's LudgerClean A cartridges are hydrophilic interaction amide cartridges (Cat # LC-A-24) which can be used for a number of applications, including clean up of:

  • 2AA/or 2AB labelled N glycans
  • VTag labelled glycopeptides
  • APTS labelled N-glycans

Each pack of LC-As contains 24 cartridges.
To order or request a quotation, contact info@ludger.com






Study of Immunoglobulin A glycosylation in pregnancy: results published in Nature Scientific Reports

Oxford, June 2016

Ludger Scientific ReportsLudger has worked in collaboration with the Department of Rheumatology, Erasmus University Medical Center, Rotterdam, The Netherlands and Leiden University Medical Centre, The Netherlands to study IgA glycosylation in pregnancy.
Serum samples were taken at different stages of pregnancy and after delivery from a cohort of 29 women. A high-throughput workflow was adopted for the simultaneous analysis of serum-derived IgA1 N- and O-glycopeptides using matrix-assisted laser/desorption ionisation Fourier transform ion cyclotron resonance (MALDI-FTICR) mass spectrometry (MS).
Pregnancy associated changes of N-glycan bisection were found to be different for IgA1 compared to IgG-Fc. This method could be used for larger patient cohorts to study IgA N- and O-glycosylation changes in pathologies.

Citation: Bondt, A. et al. Longitudinal monitoring of immunoglobulin A glycosylation during pregnancy by simultaneous MALDI-FTICR-MS analysis of N- and O-glycopeptides. Sci. Rep. 6, 27955;doi: 10.1038/srep27955 (2016).

Link to full-text publication





New Publication: Site-specific N-glycosylation analysis of HIV-1 Envelope Glycoprotein

Oxford, March 2016

Ludger Ceramide Glycanase EnzymeThe trimeric HIV type 1 (HIV-1) envelope glycoprotein (Env) is targeted by broadly neutralizing antibodies (bNAbs) produced by the immune system during infection. As part of our collaboration with Max Crispin and his group at the University of Oxford, UK, we have performed site-specific N-glycosylation analysis of the gp120 and three gp41 subunits of Env. Using MALDI-MS, LC-MS and HILIC-UPLC our results uncovered a dominance of oligomannose-type glycans and revealed a mosaic of glycan microclusters bearing under-processed glycans, especially in areas covering the gp120 outer domain and at the trimer interfaces. This information will assist in the design of Env-based vaccine immunogens.

The work has now been published in Cell Reports:

Link to full-text publication










Ludger Ceramide Glycanase Enzyme

R&D at Ludger

Oxford, February 2016

Ludger has an ongoing Research and Development programme dedicated to improving glycan analysis for biopharmaceuticals and medical glycomics. Around half of our R&D is funded by EU grants as part of collaborative research programmes.

To find out more about these exciting projects please visit our R&D page









LudgerZyme™ Ceramide Glycanase Enzyme

Oxford, February 2016

Ludger Ceramide Glycanase EnzymeGlycosphingolipids (GSLs) are implicated in the pathogenesis of various diseases including Fabry disease, Gaucher disease , Tay-Sachs disease and Sandhoff disease . The enzyme ceramide glycanase can be used for the characterisation of GSLs as it deglycosylates a variety of GSLs by cleaving the β-glycosyl linkage. Once free, the GSL glycans can be fluorescently labelled using LudgerTag labelling technology and then analysed to identify their glycosylation patterns.

Ludger’s ceramide glycanase kit (LZ-CER-HM-KIT) contains purified ceramide glycanase enzyme, buffer and and GM1 glycolipid. The kit is sufficient to deglycosylate 25 samples.

Cat # LZ-CER-HM-KIT     LudgerZyme™ Ceramide Glycanase Enzyme

To request a quote please contact info@ludger.com




Permethylation of glycans

Oxford, February 2016

ludger permethylation kitPermethylation is the most popular technique for the derivatisation of carbohydrates for MALDI-MS detection, as it enhances ionization efficiency, stabilizes the sialic acids and aids linkage analysis studies. On average one sample can take as little as 1 minute for data acquisition using MALDI-MS, making it appealing for Quality by Design (QbD) and biomarker studies. Since the conventional in-solution technique to permethylate glycans is labour intensive with long turnaround times, we have developed a microplate-based permethylation kit to give you a cost effective, high throughput and reliable method.

Our new LudgerTag kit, LT-PERMET-96, can be used to process 1-96 samples using either a manual method or an automated method that has been adapted to a liquid handling robot. LT-PERMET-96 gives excellent signal enhancement due to increased ionization efficiency and the technology has been validated according to ICH Q2 (R1) guidelines (for Analytical Validation). Intra assay repeatability CVs for relative % intensities were ≤12% for major N-glycans from human IgG with a relative % areas of ≥ 5%.The results from validation studies suggest that this permethylation technique gives data that is comparable to UHPLC from 2-AB labelled and procainamide labelled glycans.

We also sell a Pre-Permethylation Clean-up Plate (Cat# LC-PERMET-96) for enrichment of released N-glycans before the permethylation step, as well as the following permethylated human IgG glycan standards, which can be used as system suitability standards and/or as calibration standards for MALDI-TOF-MS analysis:

For more information or pricing on any of these products please contact info@ludger.com
Click here to view a poster presentation on the subject, and you can also view a full list of Ludger's recent scientific posters.





Summary of Glycan Cleanup Techniques

Oxford, November 2015

We offer a range of LudgerClean products to suit your specific need. The table below summarises the different applications.

ludger glycan cleanup table
click to enlarge



For additional technical advice, contact info@ludger.com
Orders can be sent to sales@ludger.com





Purified Glycan Standards

Oxford, September 2015

Ludger produces a comprehensive range of purified glycans, including IgG glycans, which are used as standards during the analysis of biopharmaceuticals.

Purity acceptance criteria is 85% as determined by HILIC chromatography of the 2AB labeled glycans. HPAE-PAD chromatography, MALDI mass spectrometry and NMR analysis are also performed as supporting data.

In the examples here we have given their short names and Ludger product names:

Common
Short Name
Oxford Notation Ludger Product
G0 A2 NGA2
G0F FA2 NGA2F
G1 A2G1 A2G1
G1F FA2G1 FA2G1
G2 A2G2 NA2
G2F FA2G2 NA2F

For the full list of these and other glycans (unlabeled, permethylated or labeled with 2AB, 2AA, procainamide or APTS), please visit www.ludger.com/products

If you are unsure if we have the standard you are looking for, please contact us at info@ludger.com




HIV-1 Publication in Nature Communications

Oxford, July 2015

We are delighted to announce that our collaboration with Dr Max Crispin and colleagues at the Oxford Glycobiology Institute, University of Oxford, UK, has resulted in a publication in Nature Communications. The paper is entitled "Glycan clustering stabilizes the mannose patch of HIV-1 and preserves vulnerability to broadly neutralizing antibodies".

The HIV-1 envelope glycoprotein gp120 is essential for virus entry into cells as it attaches to specific cell surface receptors. Whilst glycans on gp120 can protect the virus from antibody-mediated neutralization, a 'mannose patch' of under-processed oligomannose-type structures can also be a target for potent broadly neutralizing antibodies (bnAbs). Glycosylation site analysis on gp120 was performed using methods developed at Ludger. The contribution of individual glycosylation sites in the formation of the mannose patch is discussed in this publication, and supports the use of the mannose patch as a target for vaccine design.

http://www.ncbi.nlm.nih.gov/pubmed/26105115

Contact us for more information on Ludger's glycosylation site analysis service, info@ludger.com




Ludger V-TAG Glycopeptide Labeling and Enrichment Kit

Oxford, March 2015

We are pleased to announce the launch of a kit for the labeling and enrichment of IgG glycopeptides, enabling analysis by (U)HPLC or MALDI Mass Spectrometry.

The VTAG kit (Cat # LT-VTAG-24) is suitable for IgG subclass glycoproteins that have been digested with pronase or trypsin to release the glycopeptides. As little as 5ug IgG sample can be used. The VTAG kit labels each sample in 1 hour and enriches the sample using a solid phase extraction (SPE) device for a further hour.

This kit has been validated according to ICH guideline Q2 (R1) guidelines. Using different IgG samples and replicates of 9 for each, CVs for repeatability were typically <5%.

Cat #

LT-VTAG-24     LudgerTag™ V-Tag Glycopeptide Labeling and Enrichment Kit

To request a quote please contact info@ludger.com




Ludger Procainamide Labeling Kit

Oxford, February 2015

Procainamide labeling permits glycan identification by either mass spectrometry or UHPLC, and because of its improved ionisation efficiency compared to 2AB labeling it can permit identification of minor glycans (>1% relative peak area) by ESI-MS.

We are delighted to announce the launch of a Ludger kit for labeling glycans with procainamide (using 2PB as a reductant in place of sodium cyanoborohydride), along with a post-labeling clean up plate for the samples. This technology has been validated in house at Ludger; typical CVs for triplicate analyses were <5%.

Cat #

LT-KPROC-VP24     LudgerTag™ PROC (procainamide) Glycan Labeling Kit containing 2-picoline borane

To request a quote please contact info@ludger.com




Does Long Term Storage Affect Glycosylation of Biotherapeutics?

Oxford, October 2013

Ludger has been working in collaboration with Arecor Ltd to study the effect of storage conditions on the glycosylation of biotherapeutics. Following long term storage at elevated temperatures in Arecor’s formulation technology Arestat™, the N-glycosylation profiles of EPO and IgG were obtained. Results showed that Arecor’s technology maintains glycosylation profiles of EPO or IgG after prolonged storage at elevated temperature.

If you would like more information about this or would like to discuss how we can help you to monitor your biopharmaceutical’s glycosylation, please contact claire.morgan@ludger.com





Permethylated IgG glycan standards for MS analysis

Oxford, October 2013

Structural analysis of carbohydrates is a requirement for biopharmaceutical characterisation and may include the determination of the following features: molecular mass, composition of monosaccharides and their configurational and conformational isomers, sequence of monosaccharide residues, presence and position of branches and functional groups and interglycosidic linkages.

Mass spectrometry is a useful tool in determining the molecular weights but the sensitivity can be low especially where there are many different structures. Carbohydrates often contain carboxy, amino, sulphate, and phosphate groups and both the nature and the position of these groups on the residue and the position of this residue within the glycan may be difficult to determine. Permethylation of glycans converts hydrogen groups to methyl groups which renders the glycans hydrophobic; the conversion stabilises sialic acids and can increase signal intensity in mass measurements.

Ludger now offers a permethylated IgG glycan library to use as a system suitability standard during MS analysis. Ludger also offers a C13 version of the standard which can be used as an internal standard in the same MALDI chip spot as your C12 labelled permethylated glycans.

Cat #

CPM-C13-IGG-01     N-glycan IgG library, permethylated with heavy (13C) MeI
CPM-IGG-01   N-glycan IgG library, permethylated

To request a quote or place an order please contact info@ludger.com




LudgerSep™ C Buffer x4 concentrate for HPLC separation of sialylated, phosphorylated and sulphated glycans.

Oxford, September 2013

Many glycoproteins contain sialylated, sulphated or phosphorylated sugars, including drugs containing Mannose-6-Phosphate (Man6P). Man6P is a key targeting signal for acid hydrolase precursor proteins that are destined for transport to lysosomes and is present in therapeutic enzymes (enzyme replacement therapies) developed for treatment of lysosomal storage diseases.

Ludger has produced an ammonium formate buffer concentrate with a pH of 9 (above the pKa for phosphated glycans), which allows good separation of all anionic glycans (sialic acids, phosphorylated and sulphated sugars) using an anion exchange column.

This concentrate (Cat# LS-C-BUFFX4) simplifies the preparation of solvent for anion exchange, whilst avoiding the variations in buffer pH between batches which could lead to out-of-specification problems during glycan analysis. Simply dilute with water, add acetonitrile and use directly as an HPLC elution gradient when performing glycan analysis using the LudgerSep C3 column, Cat # LS-C3-7.5x75.

To request a quote or place an order please contact info@ludger.com




LudgerTag™ 2AB and 2AA Glycan Labelling within 2 hours

Oxford, March 2013

Ludger is delighted to announce that using the LudgerTag™ VP24 kits, glycans can be labelled with 2-aminobenzamide (2-AB) or 2-aminobenzoic acid (2-AA) within 2 hours. These kits incorporate 2-picoline borane (2-PB) reductant, a safer alternative than standard sodium cyanoborohydride. This technology has now been patented and exclusively licenced to Ludger Ltd.

The kits have been validated following ICH Q2(R1) guidelines and beta-tested in the field. Precision values were excellent, with CVs of <5% for peaks with relative areas over 5% and CVs of <8% for peak with relative areas less than 5%. The kits offer equivalent labelling efficiency as standard LudgerTag™ 2AA and 2AB labelling kits containing sodium cyanoborohydride.

Ordering information:

LudgerTag™ 2-AB labelling kit with 2PB Cat# LT-KAB-VP24

LudgerTag™ 2-AA labelling kit with 2PB Cat# LT-KAA-VP24

To request a quote or place an order please contact info@ludger.com




LudgerTag™ 2AB and 2AA Glycan Labelling Kits with Non Toxic Reductant

Oxford, November 2012

Fluorescent labelling methods for glycoprofiling of therapeutic glycoproteins are well established and aid subsequent separation and quantitation using a range of techniques. These include high performance liquid chromatography (HPLC and UHPLC), capillary electrophoresis (CE), and mass spectrometry. Of the fluorescent labels available, 2-aminobenzamide (2-AB) and 2-aminobenzoic acid (2-AA) are most widely used for oligosaccharide profiling. Existing 2-AA and 2-AB labelling kits use sodium cyanoborohydride as a reducing agent during glycan labelling. This reagent is toxic so a fume cupboard should be used during handling.

Following on from research performed by Ludger Ltd. in collaboration with Leiden University Medical Centre, The Netherlands, we have developed labelling kits containing a different and non-toxic reducing agent, 2-picoline borane (2-PB). This technology has now been patented and Ludger is delighted to offer two new kits for labelling glycans with either 2-AB (Cat# LT-KAB-VP24) or 2-AA (Cat# LT-KAA-VP24).

The kits have been validated following ICH Q2(R1) guidelines and beta-tested in the field. Precision values were excellent, with CVs of <5% for peaks with relative areas over 5% and CVs of <8% for peak with relative areas less than 5% (see Figure 1). The kits offer equivalent labelling efficiency as standard LudgerTag™ 2AA and 2AB labelling kits containing sodium cyanoborohydride (Figure 2).

Each kit comprises two premixed bottles; one containing acetic acid and DMSO solution and the other containing 2-AB or 2-AA label combined with 2PB. This makes the labelling process simpler. Labelling can also be performed within 2 hours.

Ordering information:

LudgerTag™ 2-AB labelling kit with 2PB Cat# LT-KAB-VP24

LudgerTag™ 2-AA labelling kit with 2PB Cat# LT-KAA-VP24

To request a quote or place an order please contact info@ludger.com




Quantitative Glycopeptide Standard for Accurate Monosaccharide or Sialic Acid Quantitation

Oxford, November 2012

Quantitative sialic acid or monosaccharide analysis is an important step for developers and manufacturers of biologic drugs. Regulators are putting increasing pressure on companies to perform accurate glycoprofiling on their biopharmaceuticals. These analyses fall within ICH guidelines Q6B and Q5Efor comparability studies during product development and after major manufacturing changes. Furthermore, the recent EMA monograph on monoclonal antibodies and forthcoming USP chapters <1084> and <1094>on glycosylation analysis reinforce the need to perform glycoprofiling throughout the drug life cycle. Until now the difficulty with accurately quantifying sialic acids or monosaccharides has been the lack of quantitative standards.

Ludger Ltd. has produced a purified glycopeptide standard, the first in a range of Ludger BioQuant™ quantitative standards, which can be used as an internal standard and positive control when performing sialic acid or monosaccharide analyses in house. This particular Ludger BioQuant™ Standard (Cat# BQ-GPEP-A2G2S2-10U) is a complex biantennary N-linked glycan terminating in two N-acetylneuraminic acids.

Purity of >90% has been assessed by HPLC, correct mass identity assessed by MALDI mass spectrometry. The exact amount of material has been determined by quantitative NMR (qNMR) and quantitative monosaccharide analysis (MA). Quantity values by qNMR and MA agree within 90-110%.  MA is traceable to internationally accepted references from USP and dispensed using NIST traceable labware. qNMR is traceable to a NIST SRM traceable CRM analysed to the ISO 17025 standard. A detailed certificate of analysis is given for each standard. This contains comprehensive documentation, lot-specific values, expiration date and storage information.

The BioQuant GPEP-A2G2S2 standard can be used during the sialic acid or monosaccharide release and labeling process. This will enable you to check the efficiency of glycan release, labeling and recovery and will give you confidence in the accuracy of your sialic acid or monosaccharide measurements.

To request a quote or place an order please contact info@ludger.com





Protein O-Glycosylation Analysis Review

Oxford, August 2012

Following the collaboration of Ludger with the University of Leiden, we are delighted to announce a publication in the journal Biological Chemistry.

This informative review describes the current methods available for analysis of O-glycosylation and discusses challenges that need to be met in the future. Areas covered include: analysis of released O-glycans, analysis of formerly O-glycosylated peptides (in order to give information on O-glycan attachment sites), and analysis of O-glycopeptides. Particular emphasis is given to MS fragmentation techniques such as collision-induced fragmentation, electron capture dissociation, and electron transfer dissociation.

Citation:

Gerhild Zauner, Radoslaw P. Kozak, Richard A. Gardner, Daryl L. Fernandes, André M. Deelder and Manfred Wuhrer. Biol. Chem., Vol. 393, pp. 687–708, August 2012





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