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. 

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 monoclonal antibodies using analyses optimised for each level of information you need (from intact protein to peptides/amino acids). 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.

Mass spectrometry is a very powerful technique widely used for the characterisation of biopharmaceuticals. 
Thanks to innovations in both hardware and software, this technique can now be implemented routinely. 

We presented versatile and efficient approaches for monoclonal antibodies, based on three levels of analysis: Intact protein, mAb substructures (after IdeS / FabRICATOR digestion), Peptide mapping after trypsin digestion

These were optimised in terms of sample preparation, LC separation (1D and 2D-LC), data processing / reporting

for mAbs isoform characterisation (batch-to-batch consistency, assessment of critical quality attributes, PTMs, glycosylation) and stability monitoring (mainly degradation by deamidation and oxidation). 

The performances and limitations of the different analytical approaches have been discussed, together with the advantages and drawbacks of the different LC columns and enzymes used. Particular attention will be paid to low pH trypsin digestion to minimise deamidation and oxidation for peptide mapping experiments. 
Thanks to the use of state-of-the-art analytical technologies, these reliable, sensitive and reproducible analyses can be done in a GMP regulated environment for QC and stability studies of biotech products.

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 ADCs using analyses optimised for each level of information you need. 

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

We presented efficient and optimised approaches for the characterisation of ADCs by mass spectrometry, based on different levels of analysis: 

• Intact protein for Drug-to-Antibody Ratio determination
• Substructures (after IdeS / FabRICATOR digestion) 
• 2D-LC/MS (HIC-RP) for DAR determination and identification of the different isoforms 
• Peptide mapping for determination of conjugation sites and site-occupancy 

Applications of these methods to KADCYLA^® and/or ADCETRIS^® were also presented. 

The performances and limitations of the different analytical approaches were also discussed.

Characterisation of Anti-HCP antibodies by 2D differential Gel Electrophoreses

Host-cell proteins are a major class of process-related impurities in biotherapeutics that can impact the safety and efficacy of  the product. Despite the advent of advanced analytical techniques, ELISA remains as the industry gold standard for the detection and quantification of HCPs during process development and for routine analyses.