Comprehensive Analysis of Low Abundant Mannose Glycopeptides in Peptide Mapping of Adalimumab in a Single Run

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Event Overview: http://www.chromatographyonline.com/lcgc_w/ce_methods_series

Glycosylation is a common post-translational modification (PTM) that occurs during the production of therapeutic monoclonal antibodies (mAb) and many other protein therapeutics.Reversed-phase liquid chromatography coupled to mass spectrometry (RP-LC–MS) has become a primary technique for analyzing glycosylation on antibody and protein therapeutics. However, it is challenging to chromatographically resolve some types of low abundant glycoforms based on hydrophobicity, such as high mannose‑containing glycopeptides. Capillary electrophoresis with electrospray ionization (CESI) coupled to MS provides a separation mechanism orthogonal to LC–MS based approaches, including identification and relative quantification of analytes.

This presentation provides a comparison of the peptide mapping analysis of adalimumab and its PTMs performed by both CESI-MS as well as LC–MS. Approximately 95% of the sequence was mapped with high confidence and 14 various glycopeptides were confirmed on the MS and MS/MS level using diagnostic ions. CESI-MS proved to be a strong complementary technology to ensure a high level of characterization for complex PTM analysis for the example biotherapeutic.

Key Learning Objectives:

• Achieve comprehensive characterization of various glycopeptides within a peptide mapping analysis
• Better resolve low abundant glycoforms, such as high mannose‑containing glycopeptides
• Discover orthogonal techniques to LC–MS for biotherapeutics characterization

Speaker: Dr. Zuzana Demianova, Sr. BioPharma Application Scientist, SCIEX

Time and Date: Wednesday, Aug. 26, 2020 at 11am EDT | 8am PDT | 4pm BST | 5pm CEST
On demand available after airing until Aug. 26, 2021.

Sponsor: SCIEX

Register free: http://www.chromatographyonline.com/lcgc_w/ce_methods_series