Sustainable compendial grade GMP detergent substitutes for Triton™ X-100 in bioprocessing applications

Triton™ X-100 (octyl phenol ethoxylate) has been used for decades as a detergent for viral inactivation in bioprocessing applications and the treatment of human or animal derived plasma, along with numerous cell lysis applications at laboratory scale and in cell-based biopharmaceutical production.

While the ability of Triton™ X-100 to perform in these applications has never come into question, the environmental impact of this material ultimately led to its ban in Europe as of January 2021, with the exception of existing pharmaceutical applications allowed under a public exemption or for small scale laboratory research use1.

This ban has led to significant interest in finding alternative detergents which are biodegradable, GMP compliant and pharmaceutically acceptable for use in the manufacturing of new cell-derived drug products in Europe. Alternatives to Triton™ X-100 must perform equally in regard to cell lysis and protein compatibility in order to ensure global applicability in a wide range of bioprocessing applications.

Employing xenotropic murine leukemia virus (XmuLV) and the Feline catus PG4 cell line as a model lipid-enveloped virus – host system, we compared the viral inactivation properties of over 31 different detergents belonging to 11 chemical classes. This analysis identified ten detergents that exhibited an equal log reduction in XmuLV viability compared to Triton™ X-100.

We then extended the characterization of these detergents to compare their minimum virucidal concentrations, kinetics of virus inactivation, cell lysis capabilities against three cell lines (PG4, CHO K1 and HEK293) and protein compatibility using alkaline phosphatase. Two detergents exhibited equivalent performance to Triton™ X-100 in all categories.

This study identified two chemically distinct detergent alternatives to Triton™ X-100 which are biodegradable, GMP compliant and have an established track record of pharmaceutical use and compendial compliance. These detergents represent exciting new options in the biopharmaceutical manufacturing tool kit.