LudgerLiberate Orela kit

LudgerLiberate Orela kit


  • Product Code: LL-ORELA-A2
  • Size 12 Samples

  • £519
  • Ex Tax: £519

For release of O-linked glycans from glycoprotein therapeutics.

The Ludger Liberate™ Orela Kit has been developed for O-linked deglycosylation of glycoproteins. Released glycans have free reducing terminii to allow fluorescent tagging by reductive amination. 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 significant reduction in peeling and reduces the number of sample handling steps prior to release.

 

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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 significant reduction in peeling and reduces the number of sample handling steps prior to release.

– Designed for easy, O-linked deglycosylation by QC labs.
– Straightforward and simple method.
– Released glycans have free reducing terminii suitable for fluorescent labelling prior to 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

The Orela reagent reacts at the link between the glycan and peptide backbone to liberate glycans as the glycosylamine. The acid-labile glycosylamine derivative is hydrolysed to produce the free glycan.



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 terminii 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 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:

Cat. # Item Quantity
LL-ORELAREAGENT-01 Orela Release Reagent 2 x 3ml
LL-ACETIC-50P-01 Acetic Acid Solution 2 x 3 ml
LC-CEX-H-01 LudgerClean™ CEX-H cartridges 2 x 6 cartridges
LL-REACT-01 Glass reaction vials with PTFE lined caps 2 x 6 vials
LL-COLLV-01 Glass collection vials with PTFE lined caps 2 x 6 vials