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

Warning

This publication doesn't include Institute of Computer Science. It includes Central European Institute of Technology. Official publication website can be found on muni.cz.

Authors	LOPEZ-MENDEZ Blanca BARON Bruno BRAUTIGAM Chad A JOWITT Thomas A KNAUER Stefan H UEBEL Stephan WILLIAMS Mark A SEDIVY Arthur ABIAN Olga ABREU Celeste ADAMCZYK Malgorzata BAL Wojciech BERGER Sylvie BUELL Alexander K CAROLIS Carlo DAVITER Tina FISH Alexander GARCIA-ALAI Maria GUENTHER Christian HAMACEK Josef HOLKOVÁ Jitka HOUSER Josef JOHNSON Chris KELLY Sharon LEECH Andrew MAS Caroline MATULIS Daumantas MCLAUGHLIN Stephen H MONTSERRET Roland NASREDDINE Rouba NEHME Reine NGUYEN Quyen ORTEGA-ALARCON David PEREZ Kathryn PIRC Katja PISZCZEK Grzegorz PODOBNIK Marjetka RODRIGO Natalia ROKOV-PLAVEC Jasmina SCHAEFER Susanne SHARPE Tim SOUTHALL June STAUNTON David TAVARES Pedro VANEK Ondrej WEYAND Michael WU Di
Year of publication	2021
Type	Article in Periodical
Magazine / Source	European Biophysics Journal With Biophysics Letters
MU Faculty or unit	Central European Institute of Technology
Citation
Web	https://link.springer.com/epdf/10.1007/s00249-021-01532-6?sharing_token=VFkRvz2n9bQV3JiJoWX8p_e4RwlQNchNByi7wbcMAY69Va5879vt7RWQCKX0SFb_pyW2Ir-vtv5ctoeTDSNvHVvASrK0292jB6qaMm0St-rVfV4AEkPYx9IirUcWRtcUqRai2T6zhfWw7w9KyyhJdd4fqGFbLj7zWAysEBQ0YSo%3D
Doi	http://dx.doi.org/10.1007/s00249-021-01532-6
Keywords	MST; TRIC; Benchmark; Thermophoresis; KD; Interaction
Description	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.
Related projects:	Czech Infrastructure for Integrative Structural Biology