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    Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study

    Lopez-Mendes, B. and Baron, B. and Brautigam, C.A. and Jowitt, T.A. and Knauer, S.H. and Uebel, S. and Williams, Mark and Sedivy, A. (2021) Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study. European Biophysics Journal with Biophysics letters , ISSN 0175-7571. (In Press)

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    Abstract

    Microscale thermophoresis (MST), and the closely related Temperature Related Intensity Change (TRIC), are synonyms for a recently developed measurement technique in the field of biophysics to quantify biomolecular interactions, using the (capillary-based) NanoTemper Monolith and (multiwell plate-based) Dianthus instruments. Although this technique has been extensively used within the scientific community due to its low sample consumption, ease of use, and ubiquitous applicability, MST/TRIC has not enjoyed the unambiguous acceptance from biophysicists afforded to other biophysical techniques like isothermal titration calorimetry (ITC) or surface plasmon resonance (SPR). This might be attributed to several facts, e.g., that various (not fully understood) effects are contributing to the signal, that the technique is licensed to only a single instrument developer, NanoTemper Technology, and that its reliability and reproducibility have never been tested independently and systematically. Thus, a working group of ARBRE-MOBIEU has set up a benchmark study on MST/TRIC to assess this technique as a method to characterize biomolecular interactions. Here we present the results of this study involving 32 scientific groups within Europe and two groups from the US, carrying out experiments on 40 Monolith instruments, employing a standard operation procedure and centrally prepared samples. A protein–small molecule interaction, a newly developed protein–protein interaction system and a pure dye were used as test systems. We characterized the instrument properties and evaluated instrument performance, reproducibility, the effect of different analysis tools, the influence of the experimenter during data analysis, and thus the overall reliability of this method.

    Metadata

    Item Type: Article
    Additional Information: The final publication is available at Springer via the link above. A read only version of the final published article is available at the publisher via this SharedIt link https://rdcu.be/ci8AR
    Keyword(s) / Subject(s): Biophysics, MST, TRIC, Benchmark, Thermophoresis, KD, Interaction
    School: School of Science > Biological Sciences
    Research Centres and Institutes: Structural Molecular Biology, Institute of (ISMB)
    Depositing User: Mark Williams
    Date Deposited: 12 May 2021 09:39
    Last Modified: 21 Jun 2021 02:43
    URI: https://eprints.bbk.ac.uk/id/eprint/44058

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