Profiling the Distribution of N-Glycosylation in Therapeutic Antibodies

by | Pharmaceutical

Profiling the distribution of N-Glycosylation in therapeutic antibodies using the QTRAP 6500 systemSCIEX has produced an application note demonstrating how a comprehensive MS/MS analysis of glycopeptides can be achieved by targeting the known nature of the glycosylation structures using the QTRAP® 6500 system. 

Immunoglobulin G molecules have become attractive as targeted therapeutic proteins, due to their high specificity and long circulation time. Glycosylation patterns determine the stability and bio-disposition of these recombinant protein drugs in vivo, as well as the efficacy, folding, binding affinity, specificity and pharmacokinetic properties. Therefore, a complete characterization of the biotherapeutic IgG glycosylation is desirable.

Key Advantages of Using the QTRAP® 6500 System for Quantitative Glycopeptide Profiling

  • With its 2000 m/z upper mass limit the system can sufficiently detect complex, hybrid and high mannose type containing glycopeptides.
  • High sensitivity of full scan MS/MS (EPI) acquired with low energy CID provides both carbohydrate fragment ions (mainly B/Y) and peptide fragment ions (b/y)
  • High information content MS/MS for identification of N-glycopeptides using SimGlycan® Software
  • Expanded functionality of Scheduled MRM™ Pro Algorithm enables:
    - More glycopeptides to be screened
    - More specific acquisition of MS/MS
    - Increased efficiency of targeted detection for glycopeptides without enrichment strategies.

In this application note, the feasibility of the Group triggered MIDAS™ workflow was demonstrated for the characterization and profiling of glycopeptides derived from therapeutically relevant antibodies (Trastuzumab and SiluMab) using the QTRAP® 6500 System.
The wide abundance variation in the detected glycopeptides highlights the benefit of using targeted detection on a high sensitivity QTRAP system.


Published  Jan 26, 2017

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