Access the slides of this 45 minute webinar to learn how state-of-the-art LC/MS technologies can be implemented for the efficient study of therapeutic protein glycosylation using analyses optimised for each level of information you need. 

Analytical performance of the different solutions proposed have been discussed, together with the presentation of numerous experimental results (with advantages and drawbacks) obtained during method development and optimisation. 

We presented efficient and optimized approaches for the characterisation of protein glycosylation by UPLC/mass spectrometry, based on different levels of analysis: 

• Fast and highly sensitive profiling of released N-glycans, with use of the RapiFluor-MS^® technology, 
• Specific release and profiling of O-glycans,
• Orthogonal released glycans analysis and sialylation profiling by mixed-mode chromatography, 
• Unambiguous determination of N- and O-glycosylation sites by widepore-HILIC peptide mapping, 
• High-resolution intact and subunit analysis. 

Applications of these methods to mAbs (Humira^®, Erbitux^®) and highly glycosylated proteins (Enbrel^®) will be presented. 

The performances and limitations of the different analytical approaches will be discussed. 

Although drug products issued from biotechnology have now been on the market for many years, accurate protein quantification remains a challenge. There are many colorimetric methods for protein quantification (Lowry, Bradford, BCA) but they all lack precision, and suffer from interferences and matrix effects, moerover, the accuracy is highly dependent of the protein used for calibration (...)

State-of-the art LC/MS methods applied to the characterisation of a highly-glycosylated fusion protein: Etanercept

A large number of pharmaceuticals are glycosylated proteins. An adequate glycosylation is critical for therapeutic proteins in terms of safety, bioactivity, solubility, stability, and pharmacokinetics and dynamics.