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Article Dans Une Revue European Biophysics Journal Année : 2021

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

1 UCPH - University of Copenhagen = Københavns Universitet
2 Biophysique Moléculaire (plateforme) - Molecular Biophysics (platform)
3 University of Texas Southwestern Medical Center [Dallas]
4 University of Manchester [Manchester]
5 Universität Bayreuth
6 MPIB - Max-Planck-Institut für Biochemie = Max Planck Institute of Biochemistry
7 University of London [London]
8 Vienna Biocenter Core Facilities
9 University of Zaragoza - Universidad de Zaragoza [Zaragoza]
10 UK - Univerzita Karlova [Praha, Česká republika] = Charles University [Prague, Czech Republic]
11 Warsaw University of Technology [Warsaw]
12 Institute of Biochemistry and Biophysics
13 IRS - Institut de Recherches SERVIER
14 DTU - Danmarks Tekniske Universitet = Technical University of Denmark
15 CRG - Centre for Genomic Regulation - Centre de Regulació Genòmica [Barcelona]
16 The institute of cancer research [London]
17 NKI - Netherlands Cancer Institute
18 EMBL Hamburg
19 CBM - Centre de biophysique moléculaire
20 CEITEC MU - Central European Institute of Technology [Brno]
21 LMB - MRC Laboratory of Molecular Biology [Cambridge, UK]
22 University of Glasgow
23 University of York [York, UK]
24 ISBG - Integrated Structural Biology Grenoble
25 Vilnius University [Vilnius]
26 IBCP - Institut de biologie et chimie des protéines [Lyon]
27 ICOA - Institut de Chimie Organique et Analytique
28 EMBL Heidelberg
29 National Institute of Chemistry
30 NHLBI - National Heart, Lung, and Blood Institute [Bethesda]
31 University of Zagreb
32 University of Bayreuth
33 Unibas - Université de Bâle = University of Basel = Basel Universität
34 University of Oxford
35 FCT - Faculdade de Ciências e Tecnologia [Faro]
Thomas Jowitt
Stefan Knauer
Mark Williams
Wojciech Bal
Alexander Fish
Maria Garcia-Alai
Christian Guenther
  • Fonction : Auteur
Josef Hamacek
Andrew Leech
Caroline Mas
Daumantas Matulis
Reine Nehmé
Kathryn Perez
  • Fonction : Auteur
Katja Pirc
Marjetka Podobnik
Jasmina Rokov-Plavec

Résumé

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.
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Dates et versions

hal-03481822 , version 1 (17-12-2021)

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Blanca López-Méndez, Bruno Baron, Chad Brautigam, Thomas Jowitt, Stefan Knauer, et al.. Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study. European Biophysics Journal, 2021, 50 (3-4), pp.411-427. ⟨10.1007/s00249-021-01532-6⟩. ⟨hal-03481822⟩
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