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

Lopez-Mendez, B., Baron, B., Brautigam, C. A., Jowitt, T. A., Knauer, S. H., Uebel, S., et al. (2021). Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study. European Biophysics Journal, 50, 411-427. doi:10.1007/s00249-021-01532-6.

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 Creators:
Lopez-Mendez, Blanca1, Author
Baron, Bruno1, Author
Brautigam, Chad A.1, Author
Jowitt, Thomas A.1, Author
Knauer, Stefan H.1, Author
Uebel, Stephan2, Author              
Williams, Mark A.1, Author
Sedivy, Arthur1, Author
Abian, Olga1, Author
Abreu, Celeste1, Author
Adamczyk, Malgorzata1, Author
Bal, Wojciech1, Author
Berger, Sylvie1, Author
Buell, Alexander K.1, Author
Carolis, Carlo1, Author
Daviter, Tina1, Author
Fish, Alexander1, Author
Garcia-Alai, Maria1, Author
Guenther, Christian1, Author
Hamacek, Josef1, Author
Holkova, Jitka1, AuthorHouser, Josef1, AuthorJohnson, Chris1, AuthorKelly, Sharon1, AuthorLeech, Andrew1, AuthorMas, Caroline1, AuthorMatulis, Daumantas1, AuthorMcLaughlin, Stephen H.1, AuthorMontserret, Roland1, AuthorNasreddine, Rouba1, AuthorNehme, Reine1, AuthorNguyen, Quyen1, AuthorOrtega-Alarcon, David1, AuthorPerez, Kathryn1, AuthorPirc, Katja1, AuthorPiszczek, Grzegorz1, AuthorPodobnik, Marjetka1, AuthorRodrigo, Natalia1, AuthorRokov-Plavec, Jasmina1, AuthorSchaefer, Susanne1, AuthorSharpe, Tim1, AuthorSouthall, June1, AuthorStaunton, David1, AuthorTavares, Pedro1, AuthorVanek, Ondrej1, AuthorWeyand, Michael1, AuthorWu, Di1, Author more..
Affiliations:
1external, ou_persistent22              
2Scientific Service Groups, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565170              

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Free keywords: Biophysics; MST; TRIC; Benchmark; Thermophoresis; K-D; Interaction;
 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.

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Language(s): eng - English
 Dates: 2021
 Publication Status: Published online
 Pages: 17
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Degree: -

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Title: European Biophysics Journal
Source Genre: Journal
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Publ. Info: Berlin : Springer
Pages: - Volume / Issue: 50 Sequence Number: - Start / End Page: 411 - 427 Identifier: ISSN: 0175-7571
CoNE: https://pure.mpg.de/cone/journals/resource/954925487773_1