Charge variants characterization and release assay development for co-formulated antibodies as a combination therapy

作者：Mingyan Cao, Niluka De Mel, A. G. Shannon, Meagan Prophet, Chunlei Wang, Weichen Xu, Ben Niu, Jun Kim, Methal Albarghouthi, Dengfeng Liu, Eric Meinke, Shi‐Hua Lin, Xiangyang Wang, Jihong Wang · 发表于：mAbs · 年份：2019 · DOI：10.1080/19420862.2019.1578137 · 被引用次数：37 · 研究领域：Monoclonal and Polyclonal Antibodies Research、CAR-T cell therapy research、Protein purification and stability

Combination therapy is a fast-growing strategy to maximize therapeutic benefits to patients. Co-formulation of two or more therapeutic proteins has advantages over the administration of multiple medications, including reduced medication errors and convenience for patients. Characterization of co-formulated biologics can be challenging due to the high degree of similarity in the physicochemical properties of co-formulated proteins, especially at different concentrations of individual components. We present the results of a deamidation study of one monoclonal antibody component (mAb-B) in co-formulated combination antibodies (referred to as COMBO) that contain various ratios of mAb-A and mAb-B. A single deamidation site in the complementarity-determining region of mAb-B was identified as a critical quality attribute (CQA) due to its impact on biological activity. A conventional charge-based method of monitoring mAb-B deamidation presented specificity and robustness challenges, especially when mAb-B was a minor component in the COMBO, making it unsuitable for lot release and stability testing. We developed and qualified a new, quality-control-friendly, single quadrupole Dalton mass detector (QDa)-based method to monitor site-specific deamidation. Our approach can be also used as a multi-attribute method for monitoring other quality attributes in COMBO. This analytical paradigm is applicable to the identification of CQAs in combination therapeutic molecules, and to the subsequent...