Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study.
Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study.
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DOI：
10.1007/s00249-021-01532-6
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发表时间：
2021-05
期刊：
影响因子：
--
通讯作者：
Wu D
中科院分区：
文献类型：
--
作者：
López-Méndez B;Baron B;Brautigam CA;Jowitt TA;Knauer SH;Uebel S;Williams MA;Sedivy A;Abian O;Abreu C;Adamczyk M;Bal W;Berger S;Buell AK;Carolis C;Daviter T;Fish A;Garcia-Alai M;Guenther C;Hamacek J;Holková J;Houser J;Johnson C;Kelly S;Leech A;Mas C;Matulis D;McLaughlin SH;Montserret R;Nasreddine R;Nehmé R;Nguyen Q;Ortega-Alarcón D;Perez K;Pirc K;Piszczek G;Podobnik M;Rodrigo N;Rokov-Plavec J;Schaefer S;Sharpe T;Southall J;Staunton D;Tavares P;Vanek O;Weyand M;Wu D
微尺度热电泳法(MST)和密切相关的温度相关强度变化(TIC)是生物物理学领域最近发展起来的一种测量技术的同义词，该技术使用(毛细管基)NanoTemper整体柱和(多孔板基)石榴石仪器来量化生物分子相互作用。尽管这项技术由于其低样品消耗量、易用性和普遍适用性而在科学界得到了广泛的应用，但MST/TIC并没有像其他生物物理技术(如等温滴定量热(ITC)或表面等离子体共振(SPR))那样得到生物物理学家的明确接受。这可能归因于几个事实，例如，各种(不完全了解的)效应对信号的影响，该技术仅授权给单一仪器开发商NanoTemper Technology，以及其可靠性和重复性从未经过独立和系统的测试。因此，ARBRE-MOBIEU的一个工作组已经建立了一项关于MST/TIC的基准研究，以评估该技术作为一种表征生物分子相互作用的方法。在这里，我们介绍了这项研究的结果，涉及欧洲的32个科学小组和来自美国的两个小组，在40台整体式仪器上进行实验，采用标准操作程序和集中制备的样品。以蛋白质-小分子相互作用、新开发的蛋白质-蛋白质相互作用体系和纯染料为实验体系。我们对仪器的性能进行了表征，并评估了仪器的性能、重复性、不同分析工具的效果、实验者在数据分析过程中的影响，从而评估了该方法的整体可靠性。网上版载有补充材料，可在10.1007/s00249-021-01532-6查阅。
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. The online version contains supplementary material available at 10.1007/s00249-021-01532-6.
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影响因子：
8.6
作者：
Duhr, S;Braun, D
通讯作者：
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影响因子：
3.8
作者：
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DOI：
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发表时间：
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影响因子：
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作者：
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通讯作者：
Sedivy, Arthur
DOI：
10.1016/j.chemolab.2013.07.007
发表时间：
2013-10-15
影响因子：
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作者：
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通讯作者：
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