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

J 2021

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

LOPEZ-MENDEZ, Blanca, Bruno BARON, Chad A BRAUTIGAM, Thomas A JOWITT, Stefan H KNAUER et. al.

Basic information

Original name

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

Authors

LOPEZ-MENDEZ, Blanca, Bruno BARON, Chad A BRAUTIGAM, Thomas A JOWITT, Stefan H KNAUER, Stephan UEBEL, Mark A WILLIAMS, Arthur SEDIVY, Olga ABIAN, Celeste ABREU, Malgorzata ADAMCZYK, Wojciech BAL, Sylvie BERGER, Alexander K BUELL, Carlo CAROLIS, Tina DAVITER, Alexander FISH, Maria GARCIA-ALAI, Christian GUENTHER, Josef HAMACEK, Jitka HOLKOVÁ (203 Czech Republic, belonging to the institution), Josef HOUSER (203 Czech Republic, belonging to the institution), Chris JOHNSON, Sharon KELLY, Andrew LEECH, Caroline MAS, Daumantas MATULIS, Stephen H MCLAUGHLIN, Roland MONTSERRET, Rouba NASREDDINE, Reine NEHME, Quyen NGUYEN, David ORTEGA-ALARCON, Kathryn PEREZ, Katja PIRC, Grzegorz PISZCZEK, Marjetka PODOBNIK, Natalia RODRIGO, Jasmina ROKOV-PLAVEC, Susanne SCHAEFER, Tim SHARPE, June SOUTHALL, David STAUNTON, Pedro TAVARES, Ondrej VANEK, Michael WEYAND and Di WU

Edition

European Biophysics Journal With Biophysics Letters, NEW YORK, SPRINGER, 2021, 0175-7571

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10610 Biophysics

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 2.095

RIV identification code

RIV/00216224:14740/21:00124042

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1007/s00249-021-01532-6

UT WoS

000642054100001

Keywords in English

MST; TRIC; Benchmark; Thermophoresis; KD; Interaction

Abstract

V originále

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.

Links

LM2018127, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

LOPEZ-MENDEZ, Blanca, Bruno BARON, Chad A BRAUTIGAM, Thomas A JOWITT, Stefan H KNAUER, Stephan UEBEL, Mark A WILLIAMS, Arthur SEDIVY, Olga ABIAN, Celeste ABREU, Malgorzata ADAMCZYK, Wojciech BAL, Sylvie BERGER, Alexander K BUELL, Carlo CAROLIS, Tina DAVITER, Alexander FISH, Maria GARCIA-ALAI, Christian GUENTHER, Josef HAMACEK, Jitka HOLKOVÁ, Josef HOUSER, Chris JOHNSON, Sharon KELLY, Andrew LEECH, Caroline MAS, Daumantas MATULIS, Stephen H MCLAUGHLIN, Roland MONTSERRET, Rouba NASREDDINE, Reine NEHME, Quyen NGUYEN, David ORTEGA-ALARCON, Kathryn PEREZ, Katja PIRC, Grzegorz PISZCZEK, Marjetka PODOBNIK, Natalia RODRIGO, Jasmina ROKOV-PLAVEC, Susanne SCHAEFER, Tim SHARPE, June SOUTHALL, David STAUNTON, Pedro TAVARES, Ondrej VANEK, Michael WEYAND and Di WU. Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study. European Biophysics Journal With Biophysics Letters. NEW YORK: SPRINGER, 2021, vol. 50, 3-4, p. 411-427. ISSN 0175-7571. Available from: https://dx.doi.org/10.1007/s00249-021-01532-6.

@article{1831157,
   author = {LopezandMendez, 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 GarciaandAlai, 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 Nehme, Reine and Nguyen, Quyen and OrtegaandAlarcon, David and Perez, Kathryn and Pirc, Katja and Piszczek, Grzegorz and Podobnik, Marjetka and Rodrigo, Natalia and RokovandPlavec, 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},
   article_location = {NEW YORK},
   article_number = {3-4},
   doi = {http://dx.doi.org/10.1007/s00249-021-01532-6},
   keywords = {MST; TRIC; Benchmark; Thermophoresis; KD; Interaction},
   language = {eng},
   issn = {0175-7571},
   journal = {European Biophysics Journal With Biophysics Letters},
   title = {Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study},
   url = {https://link.springer.com/epdf/10.1007/s00249-021-01532-6?sharing_token=VFkRvz2n9bQV3JiJoWX8p_e4RwlQNchNByi7wbcMAY69Va5879vt7RWQCKX0SFb_pyW2Ir-vtv5ctoeTDSNvHVvASrK0292jB6qaMm0St-rVfV4AEkPYx9IirUcWRtcUqRai2T6zhfWw7w9KyyhJdd4fqGFbLj7zWAysEBQ0YSo%3D},
   volume = {50},
   year = {2021}
}

TY  - JOUR
ID  - 1831157
AU  - 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
PY  - 2021
TI  - Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study
JF  - European Biophysics Journal With Biophysics Letters
VL  - 50
IS  - 3-4
SP  - 411-427
EP  - 411-427
PB  - SPRINGER
SN  - 01757571
KW  - MST
KW  - TRIC
KW  - Benchmark
KW  - Thermophoresis
KW  - KD
KW  - Interaction
UR  - https://link.springer.com/epdf/10.1007/s00249-021-01532-6?sharing_token=VFkRvz2n9bQV3JiJoWX8p_e4RwlQNchNByi7wbcMAY69Va5879vt7RWQCKX0SFb_pyW2Ir-vtv5ctoeTDSNvHVvASrK0292jB6qaMm0St-rVfV4AEkPYx9IirUcWRtcUqRai2T6zhfWw7w9KyyhJdd4fqGFbLj7zWAysEBQ0YSo%3D
N2  - 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.
ER  -

LOPEZ-MENDEZ, Blanca, Bruno BARON, Chad A BRAUTIGAM, Thomas A JOWITT, Stefan H KNAUER, Stephan UEBEL, Mark A WILLIAMS, Arthur SEDIVY, Olga ABIAN, Celeste ABREU, Malgorzata ADAMCZYK, Wojciech BAL, Sylvie BERGER, Alexander K BUELL, Carlo CAROLIS, Tina DAVITER, Alexander FISH, Maria GARCIA-ALAI, Christian GUENTHER, Josef HAMACEK, Jitka HOLKOVÁ, Josef HOUSER, Chris JOHNSON, Sharon KELLY, Andrew LEECH, Caroline MAS, Daumantas MATULIS, Stephen H MCLAUGHLIN, Roland MONTSERRET, Rouba NASREDDINE, Reine NEHME, Quyen NGUYEN, David ORTEGA-ALARCON, Kathryn PEREZ, Katja PIRC, Grzegorz PISZCZEK, Marjetka PODOBNIK, Natalia RODRIGO, Jasmina ROKOV-PLAVEC, Susanne SCHAEFER, Tim SHARPE, June SOUTHALL, David STAUNTON, Pedro TAVARES, Ondrej VANEK, Michael WEYAND and Di WU. Reproducibility and accuracy of microscale thermophoresis in the NanoTemper Monolith: a multi laboratory benchmark study. \textit{European Biophysics Journal With Biophysics Letters}. NEW YORK: SPRINGER, 2021, vol.~50, 3-4, p.~411-427. ISSN~0175-7571. Available from: https://dx.doi.org/10.1007/s00249-021-01532-6.

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