Glycan Labeling Cleanup - FAST - T1 Cartridges
Why Peform Post-Labeling Cleanup of Glycans? HPLC analysis of fluorescently labeled oligosaccharides is a widely used tool for biopharmaceutical glycoprofiling. In this process, glycans are released from the therapeutic glycoprotein then tagged with a fluorescent dye such as 2-AB (2-aminobenzamide) or 2-AA (2-aminobenzoic acid) by reductive amination prior to chromatography.
Excess labeling reagents such as dye and reducing agent interfere with the HPLC analysis - so need to removed without disturbing the relative proportions of the various glycan species.
S-Cartridges - Reliable but Slow One of the most reliable methods for achieving post-labeling cleanup is use of S-cartridges. These employ a hydrophilic membrane to separate the labeled glycans from non-glycan contaminants and are routinely used in the biopharma industry during glycoprofiling QC for lot release of FDA and EMA approved biopharmaceuticals. They are also the method of choice in our laboratories for detailed characterisation of drug glycosylation patterns for regulatory submissions.
S-cartridges work superbly well but they are slow and not suited for high throughput (HT) work.
SPE Alternatives to S-Cartridges - Fast and HT But Unreliable While other SPE (solid phase extraction) devices may be fast and
have higher sample throughput they may not deliver the accurate
stoichiometries and intermediate precisions of the original S-cartridge
system. Any skewing of glycosylation profiles is unacceptable
in glycoprofiling for regulatory work, comparability studies and lot
release QC. With LudgerClean S cartridges there is no selective loss of
certain glycan species such as the highly sialylated oligosaccharides.
LC-T1 Cartridges - the Reliability of S-Cartridges but Faster, Easier and High Throughout Now, after several years of work, we have developed a fast, high-throughput system for post-labeling cleanup of glycan samples with no selective loss of certain glycan species such as the highly sialylated oligosaccharides.
The Ludger T1 cartridge system is designed for drug glycoprofiling analysts who want the reliablity of S-cartridges but also want to process larger numbers of samples easily and get the results more quickly.
Cleaner Samples With Reliable Stoichiometries T1 cartridges give significant reduction in levels of excess dye whilst giving excellent glycan recovery. The cartridges have been validated following ICH Q2(R1) guidelines for cleanup of 2-AA and 2-AB labeled glycans. Precision and repeatability CVs over a range of glycan concentrations were <3%.
Clean-up can be performed in only 45 minutes.
Up to 96 samples can be processed in parallel using the Ludger-Velocity vacuum manifold.
Suitable for Samples with Neutral and Charged Glycans: Validation tests using bovine fetuin and IgG showed no selectivity between neutral and charged (sialylated) sugars.