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

López-Méndez, Blanca and Baron, Bruno and Brautigam, Chad A. and Jowitt, Thomas A. and Knauer, Stefan H. and Uebel, Stephan and Williams, Mark A. and Sedivy, Arthur and Abian, Olga and Abreu, Celeste and Adamczyk, Malgorzata and Bal, Wojciech and Berger, Sylvie and Buell, Alexander K. and Carolis, Carlo and Daviter, Tina and Fish, Alexander and Garcia-Alai, Maria and Guenther, Christian and Hamacek, Josef and Holková, Jitka and Houser, Josef and Johnson, Chris and Kelly, Sharon and Leech, Andrew and Mas, Caroline and Matulis, Daumantas and McLaughlin, Stephen H. and Montserret, Roland and Nasreddine, Rouba and Nehmé, Reine and Nguyen, Quyen and Ortega-Alarcón, David and Perez, Kathryn and Pirc, Katja and Piszczek, Grzegorz and Podobnik, Marjetka and Rodrigo, Natalia and Rokov-Plavec, Jasmina and Schaefer, Susanne and Sharpe, Tim and Southall, June and Staunton, David and Tavares, Pedro and Vanek, Ondrej and Weyand, Michael and Wu, Di. (2021) Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study. European Biophysics Journal, 50 (3-4). pp. 411-427.

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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.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Services Biozentrum > Biophysics Facility (Sharpe)
UniBasel Contributors:Sharpe, Timothy
Item Type:Article, refereed
Article Subtype:Research Article
Note:Publication type according to Uni Basel Research Database: Journal article
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Last Modified:21 Feb 2023 10:12
Deposited On:21 Feb 2023 10:12

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