Characterization of Monoclonal Antibodies Using In-Line SEC-ESI-MS

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Event Overview:

Characterization and control of monoclonal antibody (mAb) quality requires a combination of various analytical techniques during their clinical development and commercial production, with a particular focus on aggregate and fragment content. Size exclusion chromatography (SEC) remains the gold standard for determining the molecular weight (MW) distribution of mAbs expressed in mammalian cell culture. However, obtaining structural information beyond the physical size (hydrodynamic volume) typically requires the combination of SEC with mass spectrometry (MS). Over the years, native electrospray ionization (ESI) has proven to be very useful for obtaining multiply charged ions of intact proteins under near-physiological conditions. However, the analysis remains challenging and involves biomolecule-specific optimization on both the chromatography and mass spectrometry side. Here, a guide to enable users to acquire high quality intact protein mass spectra by inline SEC-ESI-MS will be presented, including the following:

• Design of MS-compatible SEC separation methods for a test mAb
• Solutions for easy method transfer from HPLC to UHPLC systems and columns
• Method considerations for ESI, including HPLC conditions and MS parameters
• Monitoring of buffer-dependent mAb conformational changes by ESI-MS

Key Learning Objectives:

• Gain knowledge on the use and optimization of volatile mass spectrometry-friendly buffers (ammonium acetate, ammonium formate, ammonium bicarbonate) in U/HPLC-MS methods for the analysis of intact mAbs
• Learn tips for the implementation of MS-compatible analytical methods using SEC for the determination of MW of biological samples with a simplified method and fast separation time
• Discover new analytical methods using SEC combined with MS for the characterization of intact mAbs, including molecular weight and their glycoform distribution
• Environmental benefits of the method are also highlighted due to no use of organic solvents

Speakers: Heidi Vitrac, Ph.D., Applications Scientist, Tosoh Bioscience LLC

Atis Chakrabarti, Ph.D., Technical Service Manager / Senior Applications Scientist, Tosoh Bioscience LLC

Times and Dates: Wednesday, May 5, 2021 at 11am EDT | 8am PDT | 4pm BST | 5pm CEST

On Demand Until May 5, 2022

Sponsor: Tosoh Bioscience

Register free: https://www.chromatographyonline.com/lcgc_w/monoclonal