Sprager_2024_J.Pharm.Sci__

Reference

Title : Identification of Acyl-Protein Thioesterase-1 as a Polysorbate-degrading Host Cell Protein in a Monoclonal Antibody Formulation Using Activity-based Protein Profiling - Sprager_2024_J.Pharm.Sci__
Author(s) : Sprager E , Moller J , Lin Y , Reisinger V , Bratkovic T , Lunder M , Vasl J , Krajnc A
Ref : J Pharm Sci , : , 2024
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.

PubMedSearch : Sprager_2024_J.Pharm.Sci__
PubMedID: 38772451
Gene_locus related to this paper: human-LYPLA1

Related information

Inhibitor ML211    ML349    ML348
Substrate Tween-80    Tween-20
Gene_locus human-LYPLA1
Family LYsophospholipase_carboxylesterase

Citations formats

Sprager E, Moller J, Lin Y, Reisinger V, Bratkovic T, Lunder M, Vasl J, Krajnc A (2024)
Identification of Acyl-Protein Thioesterase-1 as a Polysorbate-degrading Host Cell Protein in a Monoclonal Antibody Formulation Using Activity-based Protein Profiling
J Pharm Sci :

Sprager E, Moller J, Lin Y, Reisinger V, Bratkovic T, Lunder M, Vasl J, Krajnc A (2024)
J Pharm Sci :