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López-Méndez, 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 ; Hoková, Jitka ; Houser, Josef ; Johnson, Chris ; Kelly, Sharon ; Leech, Andrew ; Mas, Caroline ; Matulis, Daumantas ; McLaughlin, Stephen H. ; Montserret, Roland ; Nasreddine, Rouba ; Nehmé, Reine ; Nguyen, Quyen ; Ortega-Alarcón, David ; Perez, Kathryn ; Pirc, Katja ; Piszczek, Grzegorz ; Podobnik, Marjetka ; Rodrigo, Natalia ; Rokov-Plavec, Jasmina ; Schäfer, Susanne ; Sharpe, Tim ; Southall, June ; Staunton, David ; Tavares, Pedro ; Vanek, Ondrej ; Weyand, Michael ; Wu, Di:
Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith : a multi laboratory benchmark study.
In: European Biophysics Journal with Biophysics Letters.
Bd. 50
(2021)
.
- S. 411-427.
ISSN 1432-1017
DOI: https://doi.org/10.1007/s00249-021-01532-6
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.
Weitere Angaben
| Publikationsform: | Artikel in einer Zeitschrift |
|---|---|
| Begutachteter Beitrag: | Ja |
| Institutionen der Universität: | Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften > Fachgruppe Chemie > Ehemalige ProfessorInnen > Lehrstuhl Biopolymere - Apl. Prof. Dr. Birgitta Wöhrl Fakultäten Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften > Fachgruppe Chemie Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften > Fachgruppe Chemie > Lehrstuhl Biochemie mit Schwerpunkt Biophysikalische Chemie Fakultäten > Fakultät für Biologie, Chemie und Geowissenschaften > Fachgruppe Chemie > Ehemalige ProfessorInnen |
| Titel an der UBT entstanden: | Ja |
| Themengebiete aus DDC: | 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften; Biologie |
| Eingestellt am: | 23 Apr 2021 10:15 |
| Letzte Änderung: | 14 Aug 2023 11:28 |
| URI: | https://eref.uni-bayreuth.de/id/eprint/64905 |