LudgerClean T1 Cartridges :
For Reliable High Throughput Post-Labeling Cleanup of Biopharmaceutical Glycans
Fucose, levels of which are an important Glycosylation Critical Quality Attribute (GCQA)
for IgG1 class mAbs that use ADCC as part of their mode of action
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
Over the years we have tried many SPE (solid phase extraction) alternatives - and while these are fast and have higher sample throughput they have failed to deliver the accurate stoichiometries and intermediate precisions of the original S-cartridge system. The most common problem that we have seen with SPE alternatives is selective loss of certain glycan species such as the highly sialylated oligosaccharides. This skewing of glycosylation profiles is unacceptable in glycoprofiling for regulatory work, comparability studies and lot release QC.
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.
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™ SPE 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.
Ludger T1 cartridges are available in packs of 6 cartridges (Ludger Catalog # LC-T1-A6).
The following discounts apply when ordering multiple packs:
6-15 packs 25% discount per pack
>16 packs 50% discount per pack
- Fernandes, DL (2007) 'Future Trends in Measurement of Biopharmaceutical Glycosylation'. Presentation and discussions at the 11th CMC Strategy Forum, Washington DC, USA, January 2007
- ICH Q6B: Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological Products
- ICH Q5E: Comparability of Biotechnological/Biological Products Subject to Changes in Their Manufacturing Process
- EMA Guideline on Production and Quality Control of Monoclonal Antibodies and Related Substances (EMEA/ CHMP/ BWP/ 157653/ 2007)
- USP Chapter <1084>, Glycoprotein and Glycan Analysis – General
- USP Chapter <1095>, Glycoprotein and Glycan Analysis –Oligosaccharide Analysis
- ICH Q2(R1): Validation of Analytical Procedures: Text and Methodology
Interested in the LC-T1 Cartridges for Post-Labeling Cleanup of Glycans?
The LC-T1 cartridges and related products for using them for high-throughput glycoprofiling work are available now. Please CONTACT US for further technical or sales information.