The removal of Triton X-100 by dialysis is feasible!

Abstract

Triton X-100 has been widely used in many analytical and preparative protocols for a long time. Nevertheless, mass spectrometry, chromatographic separation, and spectrophotometric readout may be considerably hampered by this detergent due to signal suppression, complex formation, and high blank values, respectively. Additionally, Triton X-100 is not safe to remove prior to analytics. Here, microdialysis is introduced as a parallelizable, high-throughput method to clean samples from Triton X-100 with high efficacy and precision. To achieve this, we exploit the potential to considerably increase the critical micellar concentration of Triton X-100 by alteration of matrix properties. To that end, addition of several chaotropic compounds and organic solvents has been shown to increase the critical micellar concentration as well as the removal rate of the detergent. For application, matrix additives can be selected for analyte stability requirements out of a variety of compounds. Conveniently, all these additives are removable subsequently using the same microdialysis tool for downstream analytics requirements. Applicability and protocols are shown with proteomic sample preparation of purified proteins and complex protein mixtures prior to matrix-assisted laser desorption ionization (MALDI) mass spectrometry.

Triton X-100-Modified Adenosine Triphosphate-Responsive siRNA Delivery Agent for Antitumor Therapy

“…Then, the primary amine group of PEI (1.8 kDa) reacted with the activated NHS ester of Triton X-100 at an equivalent ratio of 1:6 to obtain TXPEI (Figure S1). The conjugation was purified through dialysis against 30% methanol aqueous solution for 24 h to remove the unreacted molecules and lyophilized for the following use. The conjugated product TXPEI was validated to determine the 6.32 Triton X-100 per PEI (1.8 kDa) using 1 H NMR.…”

Enhanced GAD65 production in plants using the MagnICON transient expression system: Optimization of upstream production and downstream processing

“…Several factors must be considered carefully for the development of an effective solubilization medium, including the choice and concentration of detergent used to release the target protein into solution . The introduction of surfactants into the purification process can result in the co‐extraction of a many impurities, including phenols that reduce the quality of the purified target protein by causing oxidation and that interfere with analytical methods such as protein quantification , immunological assays, isoelectric focusing, nuclear magnetic resonance spectroscopy and mass spectrometry . Furthermore, the high cost of certain surfactants can affect the economics of production.…”

Formation of Micelles by Nonionic Detergent Molecules Leads to the Breakthrough Peak in Reversed-Phase Ultraperformance Liquid Chromatography (UPLC)

“…Although procedures have been recommended to mitigate solvent strength mismatch issues, 21 our results suggest that novel strategies must be pursued to eliminate surfactants from water-based solvent systems. For instance, using high-quality commercial water not produced by a filtration system or purifying water by anion exchange, 22 gel filtration, or dialysis 23 could be potential solutions. Installing a guard column in the aqueous solvent supply line before it enters the injection loop will effectively minimize the effects of detergent accumulation on the analytical column.…”