CE methods for charge variant analysis of mAbs and complex format biotherapeutics.

Autor: Meudt M; Analytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany.; Institute of Pharmaceutical Biotechnology, Ulm University, Ulm, Germany., Pannek M; Analytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany.; Rentschler Biopharma SE, Laupheim, Germany., Glogowski N; Analytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany., Higel F; Global CMC Experts NBE, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany., Thanisch K; Analytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany., Knape MJ; Analytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany.
Jazyk: angličtina
Zdroj: Electrophoresis [Electrophoresis] 2024 Aug; Vol. 45 (15-16), pp. 1295-1306. Date of Electronic Publication: 2024 Jan 17.
DOI: 10.1002/elps.202300170
Abstrakt: Charge heterogeneity analysis of monoclonal antibodies (mAbs) and complex formats, such as bispecifics, is crucial for therapeutic applications. In this study, we developed two capillary electrophoresis (CE)-based methods, capillary zone electrophoresis (CZE) and imaged capillary isoelectric focusing (iCIEF), for analyzing a broad spectrum of mAbs and complex mAb formats. For CZE, we introduced a new buffer system and optimized the background electrolyte (BGE) with an alternative dynamic coating agent and a superior polymeric additive. The pH of the BGE was increased, leading to enhanced resolution of high pI and complex format mAbs. In iCIEF, we identified an ampholyte combination offering a highly linear pH gradient and covering a suitable pH range. We also investigated alternatives to denaturing stabilizers and found that non-detergent sulfobetaine 195 exhibited excellent properties for iCIEF applications. These optimized methods provide a framework for the charge heterogeneity analysis of therapeutic mAbs and complex formats.
(© 2024 Boehringer Ingelheim Pharma GmbH & Co KG. ELECTROPHORESIS published by Wiley‐VCH GmbH.)
Databáze: MEDLINE
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