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Facile measurement of protein stability and folding kinetics using a nano differential scanning fluorimeter

Chattopadhyay, Gopinath and Varadarajan, Raghavan (2019) Facile measurement of protein stability and folding kinetics using a nano differential scanning fluorimeter. In: PROTEIN SCIENCE, 28 (6). pp. 1127-1134.

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Official URL: https://doi.org/10.1002/pro.3622

Abstract

With advancements in high-throughput generation of phenotypic data on mutant proteins, it has become important to individually characterize different proteins or their variants rapidly and with minimal sample consumption. We have made use of a nano differential scanning fluorimetric device, from NanoTemper technologies, to rapidly carry out isothermal chemical denaturation and measure folding/unfolding kinetics of proteins and compared these to corresponding data obtained from conventional spectrofluorimetry. We show that using sample volumes 10-50-fold lower than with conventional fluorimetric techniques, one can rapidly and accurately measure thermodynamic and kinetic stability, as well as folding/unfolding kinetics. This method also facilitates characterization of proteins that are difficult to express and purify.

Item Type: Journal Article
Additional Information: copyright for this article belongs to PROTEIN SCIENCE
Keywords: nano differential scanning fluorimeter; isothermal denaturation; thermodynamic stability; refolding unfolding kinetics
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Depositing User: Id for Latest eprints
Date Deposited: 27 Jun 2019 14:23
Last Modified: 27 Jun 2019 14:23
URI: http://eprints.iisc.ac.in/id/eprint/63045

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