EXCLUSIVE WHITEPAPER: Next-generation Triton™ X-100 replacements for pharmaceutical bioprocessing

Figure 1 VI by three competitors compared to Virodex™ TXR-1 and TXR-2 and Triton™ X-100. All detergents were tested at 22 °C at a concentration of 0.1 % (v/v), unless indicated otherwise. The dotted red line indicates the log-reduction factor (LRF) at which no additional virus can be detected. 

Virodex™ and biologic product integrity 

The Virodex™ detergents are non-denaturing detergents that do not affect the structure of proteins, making them ideal reagents for processes that generate protein-based therapeutics or non-enveloped viral vectors used in CGT products. 

Virodex™ TXR-1 and TXR-2 do not affect the enzymatic activity of secreted embryonic alkaline phosphate (SEAP) activity at concentrations up to 2.5 %, and their protein compatibility is equivalent to that of Triton™ X-100. Good protein compatibility is consistent with the non-ionic nature of the Virodex™ detergents. The anionic detergent sodium dodecyl sulphate (SDS) shows strong enzyme inactivation at concentrations > 0.3 % (Fig.2).

Figure 2 Activity of SEAP after detergent treatment: Expression of SEAP by HEK-293T cells was induced by treatment with tumor necrosis factor alpha (TNFa) and 100 % activity was defined as SEAP activity in untreated controls (pink dashed line). Base-line SEAP activity (0 %) was determined using cells that were not induced with TNFa (pink dotted line). Following TNFa activation (24 h) cells were treated with detergents for 2 h at 37 °C and then an aliquot of the cell culture medium was mixed with HEK-Blue Detection medium (Invivogen) and incubated for 24 h at 37 °C and 5 % CO2.

Compatibility with traditional Protein A chromatography for detergent removal from final drug product  

Analytical detection methods were utilised to assess the affinity of the Virodex products to protein A resin, wherein no binding was seen demonstrating the ease of depletion in standard downstream processing (Fig.3).

Figure 3 Protein A column affinity of Virodex™ TXR-1 and TXR-2. Protein A resin affinity was assessed using MabCaptureC™; MiniChrom columns (ThermoFisher₁). Virodex™ detergents in chromatography eluates was measured by HPLC-ELSD. The limit of quantification was 5 ppm for Virodex™ TXR-1 and 10 ppm for Virodex™ TXR-2. One column volume (CV) was 1 mL. 

Choose Virodex™: Effective, sustainable, REACH compliant alternatives to Triton™ X-100 

 

1. https://www.fishersci.fi/shop/products/mabcapturec-minichrom-column/30019344 

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