2020
DOI: 10.1007/s12551-020-00619-2
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Theory and applications of differential scanning fluorimetry in early-stage drug discovery

Abstract: Differential scanning fluorimetry (DSF) is an accessible, rapid, and economical biophysical technique that has seen many applications over the years, ranging from protein folding state detection to the identification of ligands that bind to the target protein. In this review, we discuss the theory, applications, and limitations of DSF, including the latest applications of DSF by ourselves and other researchers. We show that DSF is a powerful high-throughput tool in early drug discovery efforts. We place DSF in… Show more

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Cited by 175 publications
(164 citation statements)
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“…Because DSF relies on the use of dye fluorescence as a proxy for protein unfolding, identifying and minimizing sources of protein-independent fluorescence is a critical step. Indeed, the fluorescence of SYPRO Orange is known to be sensitive to excipients that are common in biological buffers, such as glycerol, detergents, lipids and EDTA 34,43 . Here, we focus on two especially pernicious and common sources of protein-independent fluorescence: dye binding to colloidal aggregates and dye binding to the plastic used in manufacture of some microtiter plates.…”
Section: Include No-protein Controls For Every Conditionmentioning
confidence: 99%
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“…Because DSF relies on the use of dye fluorescence as a proxy for protein unfolding, identifying and minimizing sources of protein-independent fluorescence is a critical step. Indeed, the fluorescence of SYPRO Orange is known to be sensitive to excipients that are common in biological buffers, such as glycerol, detergents, lipids and EDTA 34,43 . Here, we focus on two especially pernicious and common sources of protein-independent fluorescence: dye binding to colloidal aggregates and dye binding to the plastic used in manufacture of some microtiter plates.…”
Section: Include No-protein Controls For Every Conditionmentioning
confidence: 99%
“…The Tma is then calculated as the midpoint of the resulting fluorescence versus temperature curve, and higher Tmas suggest a more stable native protein. Because Tmas are broadly sensitive to changes in the biochemical state of the native protein, such as the binding of ligands [1][2][3][4][5][6][7] , mutations 8,9 , and proteinprotein interactions 10,11 , DSF is widely used to monitor diverse biochemical events via the associated change in Tma (∆Tma) 12,13 .…”
Section: Introductionmentioning
confidence: 99%
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“…Jumping up one level in the scale of observation, the next review article by Gao et al also deals with the subject of protein unfolding, but in this case, their focus is on the technique of differential scanning fluorimetry (DSF), one of the most commonly used procedures for recording a temperature-induced unfolding transition (Gao et al 2020). By reviewing examples of DSF-based studies utilizing both the extrinsic and intrinsic fluorescence modes, the authors outline experimental considerations and contrast the advantages of DSF against other experimental procedures for monitoring protein stability.…”
Section: Description Of Issue Contentsmentioning
confidence: 99%