Glycan Analysis Services

Role and importance of glycosylation

Protein glycosylation can play an important role in bioactivity, stability, biological half-life and immunogenicity of a biopharmaceutical. Glycosylation is a post-translational modification and, unlike transcription, is a non-template-driven enzymatic modification process, so glycosylation can change with alterations to production conditions. Glycosylation 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.

Analysis of glycosylation is important not only in biopharmaceutical research but also in clinical and biological research where changes in glycosylation have been associated with many states of health and disease providing prognostic and diagnostic information.

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Sample types

Ludger's Glycan Analysis Team has many years of expertise with analysing glycosylation (including N- and O- glycosylation) from 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
• COVID-19 patient samples (e.g. plasma, tissues)
• SARS-CoV-2 infected cell lines

Ludger has been offering custom analytical services to suit your individual requirements since 1999.

We listen and respond with dedicated tailored solutions. Our data and customised reports are used:

• in QbD studies and early stages of drug development
• in process optimisation and production scale-up
• in comparability studies (biosimilars, biobetters)
• to support regulatory submissions
• for lot release of drug batches during biomanufacturing
• in research & method development

Our philosophy is to work with you in a partnership to ensure successful delivery of the information you need.

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Regulatory requirements & Ludger's approach

EMA guideline on development, production, characterisation and specification for monoclonal antibodies and related products states:

"The carbohydrate content (neutral sugars, amino sugars and sialic acids) should be determined. In addition, the structure of the carbohydrate chains, the oligosaccharide pattern (antennary profile), the glycosylation site(s) and occupancy should be analysed.

Typically, monoclonal antibodies have one N-glycosylation site on each heavy chain located in the Fc region. The light chain is usually not glycosylated. However, additional glycosylation site(s) in the heavy chains may occur, and thus their presence or absence should be confirmed. Glycan structures should be characterised, and particular attention should be paid to their degree of mannosylation, galactosylation, fucosylation and sialylation. The distribution of the main glycan structures present (often G0, G1 and G2) should be determined.

Higher-order structure of the monoclonal antibody should be characterised by appropriate physicochemical methodologies."

ICH Q6B guideline states:

"For glycoproteins, the carbohydrate content (neutral sugars, amino sugars, and sialic acids) is determined. In addition, the structure of the carbohydrate chains, the oligosaccharide pattern (antennary profile), and the glycosylation site(s) of the polypeptide chain are analyzed, to the extent possible."

At Ludger we use a systematic approach aligned with current regulatory guidelines to support drug developers and researchers. Our system has three broad steps:

1. Specification of Glycosylation Critical Quality Attributes (GCQAs) (i.e. those glycosylation parameters that most influence the drug product's safety and efficacy profiles).
2. Implementation of appropriate, affordable glycoprofiling modules to measure the GCQAs throughout the drug's life cycle.
3. Interpretation of the glycoprofiling data and taking appropriate action if the product falls out of specification (OOS) or trends towards OOS.

Regulatory requirements	Ludger Glycan Analysis Services
Quantitative Monosaccharide Analysis	✓
Quantitative Sialic Acid Analysis	✓
Glycan Profiling Analysis	✓
Glycan Antennary Profiling Analysis	✓
Linkage Analysis	✓
Detailed Glycan Characterisation	✓
Glycosylation Site Analysis	✓

GCQAs	Ludger Glycan Analysis Services
Degree of Mannosylation	✓
Degree of Galactosylation	✓
Distribution of G0, G1 and G2	✓
Degree of Fucosylation	✓
Degree of Sialylation/Charge distribution	✓
Human vs. Non-Human Sialic Acids	✓
Detection and Relative Quantitation of N-glcayns Containing the Galα1-3Gal Epitope	✓
Compliance documentation for GMP	✓

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Ludger's glycoprofiling programmes:

Hover over each module to zoom (or pinch to zoom on touch devices), and click for more information/to visit webpages for each module.

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Contacts