New Croda product range Virodex™: Replacements to Triton™ X-100

Triton™ X-100 is used widely in the biopharmaceutical industry for virus inactivation and cell lysis applications. While the performance of Triton™ X-100 cannot be challenged, the detergent is associated with a number of disadvantages, with the product degrading to endocrine disrupting by-products, making it an aquatic reproductive toxin.It is also included on the European Chemical Agency’s candidate list of substances of very high concern of the REACH regulation (Registration, Evaluation, Authorization and Restriction of Chemicals).

As a result, Triton™ X-100 alternatives have been developed, including our newly launched product range, Virodex™, addressing the industry need to find a suitable replacement for viral inactivation and/or cell lysis in their bioprocessing applications.

Watch the video: Sustainable compendial grade GMP detergent substitutes for Triton™ X-100

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

Abstract

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 TritonTM
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 TritonTM 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 TritonTM 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.

Introduction

TritonTM X-100 is a 4-(1,1,3,3-tetramethylbutyl)-phenyl ethoxylate detergent that is used widely in the
biopharmaceutical industry for virus inactivation and cell lysis applications. TritonTM X-100 degrades to endocrine disrupting by-products making it an aquatic reproductive toxin2. This led to its inclusion in the European Chemical Agency’s candidate list of substances of very high concern of the Registration, Evaluation, Authorization and Restriction of Chemicals (REACH) regulation3. TritonTM X-100 was banned for use in the EU on Jan 4, 2021 with a few exemptions, requiring biopharmaceutical manufacturers to find alternative detergents. Replacements for TritonTM X-100 should be eco-friendly and functionally equivalent or superior to TritonTM X-100. Alternatives to TritonTM X-100 have been developed4 but there is room in the market to explore detergents from Croda’s portfoliothat can act as suitable replacements.

To identify Croda detergents with equivalent or superior performance to TritonTM X-100, 31 detergents belonging to 11 structural classes were evaluated for their virus inactivation properties and compared to TritonTM X-100 and three other industry benchmarks. To profile the virus inactivation activity of the detergents, assays utilizing XMuLV were developed as this virus is commonly used as a model for human retroviruses5. This study was conducted in two phases. In the first phase we screened 31 detergents to identify the highest performing detergents. In the second phase we compared two prioritized detergents to industry benchmarks. The results of this work identified Virodex TXR-1 and TXR-2 as exciting new products for virus inactivation and cell lysis applications in the bioprocessing and biopharmaceutical industries.

See the full technical brochure on “Triton™ X-100” here

(click the picture to download the brochure)

Source: Croda technical brochure “Triton™ X-100“

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