Identification and characterization of a Triton X‐100 replacement for virus inactivation

Abstract

Triton X-100 detergent treatment is a robust enveloped virus inactivation unit operation included in biopharmaceutical manufacturing processes. However, the European Commission officially placed Triton X-100 on the Annex XIV authorization list in 2017 because a degradation product of Triton X-100, 4-(1,1,3,3-tetramethylbutyl) phenol (also known as 4-tert-octylphenol), is considered to have harmful endocrine disrupting activities. As a result, the use of Triton X-100 in the European Economic Area (EEA) would not be allowed unless an ECHA issued authorization was granted after the sunset date of January 4, 2021. This has prompted biopharmaceutical manufacturers to search for novel, environment-friendly alternative detergents for enveloped virus inactivation. In this study, we report the identification of such a novel detergent, Simulsol SL 11W. Simulsol SL 11W is an undecyl glycoside surfactant produced from glucose and C11 fatty alcohol. We report here that Simulsol SL 11W was able to effectively inactive enveloped viruses, such as xenotropic murine leukemia virus (XMuLV) and pseudorabies virus (PRV). By using XMuLV as a representative enveloped virus, the influence of various parameters on the effectiveness of virus inactivation was evaluated. Virus inactivation by Simulsol SL 11W was effective across different clarified bioreactor harvests at broad concentrations, pH, and temperature ranges. Simulsol SL 11W concentration, temperature of inactivation, and treatment time were identified as critical process parameters for virus inactivation. Removal of Simulsol SL 11W was readily achieved by Protein A chromatography and product quality was not affected by detergent treatment. Taken together, these results have shown the potential of Simulsol SL 11W as a desirable alternative to Triton X-100 for enveloped virus inactivation that could be readily implemented into biopharmaceutical manufacturing processes.

Supported Lipid Bilayer Platform for Characterizing the Membrane-Disruptive Behaviors of Triton X-100 and Potential Detergent Replacements

“…Accordingly, to test detergent candidates, the major focus has been on treating selected membrane-enveloped viruses with different detergent concentrations under varying environmental conditions and exposure times and measuring the resulting effects on viral infectivity. This experimental approach has led to identifying several detergents with potentially suitable virus-inactivating properties, including the following two compounds: (1) Simulsol SL 11W (SL-11W), which is a commercially available glycoside surfactant that consists of an 11-carbon long, fatty alcohol joined together with a glucose molecule [ 12 ]; and (2) Nereid, which is a recently developed, phenol-free detergent that is structurally similar to TX-100 and not yet commercially available [ 13 ]. Depending on the study, the detergent candidates have either been tested alone or in direct comparison with TX-100, and the quantitative basis for performance evaluation has been the magnitude of the drop in viral infectivity and the corresponding inactivation kinetics.…”

“…For example, TX-100 inhibits membrane-enveloped viruses [ 15 , 16 , 17 , 18 ] and causes membrane solubilization [ 19 , 20 ], while SL-11W has also been reported to inhibit membrane-enveloped viruses and is speculated, but not proven, to cause membrane solubilization [ 12 ]. It is important to investigate such mechanistic possibilities while more broadly developing experimental capabilities that can evaluate prospectively the membrane-disruptive properties of detergent candidates to replace TX-100.…”

Identification of compendial nonionic detergents for the replacement of Triton X‐100 in bioprocessing

“…9 Thus, the industry has been searching for suitable replacements for Triton X-100. [10][11][12][13] Compendial monographs are documents maintained by private non-governmental standard setting bodies such as the United States Pharmacopeia (USP) or the European Pharmacopeia (PhEur). These documents contain chemical and testing information for many pharmaceutical inactive and active ingredients.…”

Integrated autolysis, DNA hydrolysis and precipitation enables an improved bioprocess for Q-Griffithsin, a broad-spectrum antiviral and clinical-stage anti-COVID-19 candidate

“…Next, we desired to determine whether the existing autolysis method could be adapted to better fit a large-scale and low-cost bioprocess. Specifically, we sought to avoid its reliance on a freeze-thaw cycle, which is not amenable to large-scale processing, and on Triton X-100, which is no longer generally allowed by European drug regulators [24] . We reasoned that the freeze-thaw cycle could be avoided if elevated temperatures caused a similar membrane disruption without deactivating the lysozyme and nuclease of DLF_R004.…”