Krajnc, Aleksander, Lin, Yu-Hsien, Moeller, Jens Reinhold, Reisinger, Veronika Bettina, Vasl, Jozica and Krajnc, Aleksander (2024) Identification of Acyl-Protein Thioesterase-1 as a Polysorbate-Degrading Host Cell Protein in a Monoclonal Antibody Formulation Using Activity-Based Protein Profiling. Journal of pharmaceutical sciences. ISSN 00223549

Abstract

Polysorbate (PS) degradation in monoclonal antibody (mAb) formulations poses a significant challenge in the biopharmaceutical industry. PS maintains protein stability during drug product's shelf life but is vulnerable to breakdown by low-abundance residual host cell proteins (HCPs), particularly hydrolytic enzymes such as lipases and esterases. In this study, we used activity-based protein profiling (ABPP) coupled with mass spectrometry to identify acyl-protein thioesterase-1 (APT-1) as a polysorbate-degrading HCP in one case of mAb formulation with stability problems. We validated the role of APT1 by matching the polysorbate degradation fingerprint in the mAb formulation with that of a recombinant APT1 protein. Furthermore, we found an agreement between APT1 levels and PS degradation rates in the mAb formulation, and we successfully halted PS degradation using APT1-specific inhibitors ML348 and ML211. APT1 was found to co-purify with a specific mAb via hitchhiking mechanism. Our work provides a streamlined approach to identifying critical HCPs in PS degradation, supporting quality-by-design principles in pharmaceutical development.

Item Type:	Article
Keywords:	Antibody drug(s) Enzyme(s) Protein formulation(s) Hydrolysis Liquid Chromatography-Mass spectrometry (LC-MS) Monoclonal antibody(s) Surfactant(s)
Date Deposited:	14 Jun 2024 00:46
Last Modified:	14 Jun 2024 00:46
URI:	https://oak.novartis.com/id/eprint/51996