LudgerLiberate Orela kit

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  Product guide

Product specification

The kit includes reagents and materials for O-linked deglycosylation of up to 12 glycoprotein samples analysed in parallel or two sets of 6 samples, typically up to 1 mg of glycoprotein per sample. This method of O-glycan release demonstrates a significant reduction in peeling and reduces the number of sample handling steps before release.

– Designed for easy, O-linked deglycosylation by QC labs.
– Straightforward method.
– Released glycans have free reducing termini suitable for fluorescent labelling before analysis by HPLC and MS.
– Fluorescent labelling allows relative quantitation of glycan species by HPLC.
– Faster and lower cost than hydrazinolysis (LL-HYDRAZ-A2).
– No special handling techniques.

The Orela Reaction

Liberation of the glycans as the hydroxyl derivatives

The Orela reagent reacts as a base on the terminal monosaccharide, attached to the serine or threonine, to cleave the covalent bond between the glycan and peptide backbone to liberate glycans with a terminal cyclic monosaccharide with a hydroxyl functional group.

Application: For release of O-linked glycans from glycoprotein therapeutics

Description: The kit contains reagents for the release of O-linked glycans from glycoprotein biopharmaceuticals. Released glycans have free reducing termini to allow fluorescent tagging by reductive amination.

Number of Samples: The kit contains reagents and materials for up to 12 glycoprotein samples analysed in parallel or two sets of 6 samples.

Amount of Sample: Typically, up to 1 mg of glycoprotein per sample.

Suitable Samples: Biopharmaceutical glycoproteins.

Storage: Store refrigerated at 4 to 10 °C in the dark. If you have limited cold storage space, store the LudgerClean™ CEX-H cartridges (Cat # LC-CEX-H-01) at 4°C and the rest of the kit at room temperature. Protect from sources of heat, light, and moisture. The reagents are stable for at least 18 months from date of manufacture.

Shipping: The product should be shipped at ambient temperature.

For research use only. Not for human or drug use

Additional Reagents and Equipment Required 

• Pure water: resistivity 18 MΩ-cm, particle free (>0.22 μm), TOC <10 ppb
• Dialysis membranes, PD10 columns, centrifugal concentrators§ or similar for removal of salts and detergents from your glycoprotein samples*
• Glove box with dry, oxygen-free inert gas (e.g. nitrogen or argon) for handling anhydrous hydrazine. As an alternative to a glove box, hydrazinolysis can be performed in a dry fume hood with an inverted funnel applying a blanket of dry argon.
• Syringe (glass or PTFE) to transfer anhydrous hydrazine – e.g. 1 ml Hamilton or SGE glass syringe for liquids with Teflon tipped plunger and stainless steel or PTFE needle. Do not use plastic syringes.
• Heating block, oven, or similar dry heater (a water bath cannot be used) that can be set at between 40°C and 100°C
• Centrifugal evaporator (e.g.ThermoSavant SpeedVac®, Heto or GeneVac®). If using the ThermoSavant SpeedVac® we recommend the use of the Thermo Savant RH32-13 Rotor.
• Pipettes 

This kit contains the following materials and reagents: