Julia Baek1, Rosa Viner2, Terry Zhang2, Eugen Damoc3, Shanhua Lin1
ASMS 2019 Poster Purpose: To optimize a volatile pH gradient mobile phase system for native mass spectrometry analysis of monoclonal antibody charge variants using weak cation exchange (WCX) chromatography.
Methods: Weak cation exchange chromatography using volatile pH gradient directly coupled to the Orbitrap mass spectrometer for charge variant analysis of four therapeutic monoclonal antibodies (Vedolizumab, Secukinumab, Pertuzumab and Trastuzumab).
Results: Volatile pH gradient buffers separated multiple charge variants of four mAbs with different pI values. Lysine variants were separated from the main product peak in the WCX separation and glycosylation forms of different lysine variant were identified using ReSpect deconvolution and Sliding Window analysis in BioPharma Finder 3.1. In addition, deamidation was detected on Secukinumab.
1Thermo Fisher Scientific 11228 Titan Way, Sunnyvale, CA 94085, USA; 2355 River Oaks Parkway, San Jose, CA 95134, USA; 3Hanna-Kunath-Str. 11, 28199 Bremen, Germany