Profiling Active Enzymes for Polysorbate Degradation in Biotherapeutics by Activity-Based Protein Profiling

Polysorbate is widely used to maintain stability of biotherapeutic proteins in pharmaceutical formulation development. Degradation polysorbate can lead particle formation drug products, which a major quality concern and potential patient risk factor. Enzymatic activity from residual host cell enzymes such as lipases esterases plays role for degradation. Their high activity, often at very low concentration, constitutes analytical challenge the biopharmaceutical industry. In this study, we evaluated optimized activity-based protein profiling (ABPP) approach identify active responsible Using an chemical probe, established first global profile serine hydrolases harvested culture fluid (HCCF) monoclonal antibodies (mAbs) production two Chinese hamster ovary (CHO) lines. A total eight known were identified by ABPP with enzyme information, while only five traditional abundance-based proteomics (TABP) approach. Interestingly, phospholipase B-like 2 (PLBL2), well-known problematic HCP was not found be process-intermediates different mAbs. proof-of-concept study downstream samples, A2 group VII (PLA2G7) confirmed contribute polysorbate-80 degradation time. The ABBP approved able low-abundance fills gap between lipase abundance enables more meaningful investigations