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

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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.

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Abbreviations

BSA:

Bovine serum albumin

CA:

Carbonic anhydrase

CMC:

Critical micellar concentration

CV:

Coefficient of variation

DLS:

Dynamic light scattering

DMSO:

Dimethylsulfoxide

DTT:

Dithiothreitol

GHC:

Guanidine hydrochloride

H33342:

Hoechst dye bisbenzimide trihydrochloride

HCl:

Hydrochloride

LOQ:

Limit of quantification

MALDI-TOF:

Matrix-assisted laser desorption ionization–time of flight

MD100:

Xpress Micro Dialyzer (for volumes of up to 100 μl of sample)

MWCO:

Molecular weight cutoff

NaI:

Sodium iodide

NaSCN:

Sodium thiocyanate

NH4HCO3 :

Ammonium bicarbonate

SD:

Standard deviation

TB:

Tris buffer (containing 100 mM Tris/HCl, pH 7.4)

TEB:

Triethanolamine buffer (containing 100 mM triethanolamine hydrochloride/HCl and 0.02 % (w/v) sodium azide, pH 7.4)

TFA:

Trifluoroacetic acid

Tris:

Tris(hydroxymethyl)aminomethane

TX:

Triton X-100

α-CD:

α-Cyclodextrin

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Acknowledgments

We thank Dr. Holger Wondraczek and Thomas Elschner (Institute for Organic Chemistry and Macromolecular Chemistry, Friedrich Schiller University Jena) for giving us the opportunity to measure on Zetasizer Nano ZS and their kind help, Sindy Wendler and Dr. Rita Büchler (Institute of Biochemistry I) for their technical support, and Dr. Rolf Nitzsche (Malvern Instruments) for discussing our results from DLS measurements. For language editing, Laura McMillan (Institute of Biochemistry I) is gratefully acknowledged. The financial support of the Federal Ministry for Economic Affairs and Energy and the Central Innovation Program SME (KF 2937601FR1) and of the DFG Collaborative Research Center/Transregio 124 – FungiNet (project Z2) is gratefully acknowledged.

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Authors and Affiliations

  1. Institute of Biochemistry I, Jena University Hospital, Nonnenplan 2, 07743, Jena, Germany

    Stefan Opitz, Franziska Hannika & Heidrun Rhode

  2. Department of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Hans Knöll Institute Jena, Adolf-Reichwein-Straße 23, 07745, Jena, Germany

    Thomas Krüger

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  1. Stefan Opitz
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  2. Franziska Hannika
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Correspondence to Heidrun Rhode.

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Opitz, S., Hannika, F., Krüger, T. et al. The removal of Triton X-100 by dialysis is feasible!. Anal Bioanal Chem 407, 1107–1118 (2015). https://doi.org/10.1007/s00216-014-8333-3

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  • DOI: https://doi.org/10.1007/s00216-014-8333-3

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