Exploring the design space for Triton X‐100 substitutes in viral inactivation applications

Abstract The urgent need to replace the European‐prohibited Triton X‐100 in biomanufacturing has been hindered by insufficient data on alternative detergents' minimum effective concentrations (MECs) and process robustness in viral inactivation. This study makes systematic research including: (1) Establishment of MECs for novel Triton X‐100 substitutes (TXR‐1/VIS/13‐S9/C16) achieving effective inactivation of Xenotropic murine leukemia virus and Pseudorabies virus (log 10 reduction factor >4) across diverse CHO harvest fluids; (2) Demonstration of broad‐spectrum efficacy against various viruses, with TXR‐1/VIS/13‐S9 maintaining effective inactivation for Bovine viral diarrhea virus , Vesicular stomatitis virus , Baculovirus , and Herpes simplex virus type 1; (3) Identification of PS20's material‐dependent inactivation dynamics, establishing standalone parameters (4 h at 37°C) that achieve equivalent viral inactivation to traditional tri(n‐butyl)phosphate ‐combined methods without requiring lipase activity—a paradigm shift in detergent application. Crucially, process optimization revealed that extending exposure time (1–4 h) enhanced PS20/PS80 efficacy more effectively than two fold concentration increases, providing cost‐effective solutions. These findings deliver broader design spaces for implementing eco‐friendly detergents while ensuring compliance with EMA/ICH viral safety standards.