DSF Guided Refolding As A Novel Method Of Protein Production

Amadeo B. Biter; Andres H. de la Peña; Roopa Thapar; Jean Z. Lin; Kevin J. Phillips

doi:10.1038/srep18906

journal article Open Access Jan 19, 2016

DSF Guided Refolding As A Novel Method Of Protein Production

Amadeo B. Biter Andres H. de la Peña

Roopa Thapar Jean Z. Lin Kevin J. Phillips

Scientific Reports Vol. 6 No. 1 · Springer Science and Business Media LLC

View at Publisher Save 10.1038/srep18906

Abstract

AbstractThe Anfinsen hypothesis, the demonstration of which led to the Nobel prize in Chemistry, posits that all information required to determine a proteins’ three dimensional structure is contained within its amino acid sequence. This suggests that it should be possible, in theory, to fold any protein in vitro. In practice, however, protein production by refolding is challenging because suitable refolding conditions must be empirically determined for each protein and can be painstaking. Here we demonstrate, using a variety of proteins, that differential scanning fluorimetry (DSF) can be used to determine and optimize conditions that favor proper protein folding in a rapid and high-throughput fashion. The resulting method, which we deem DSF guided refolding (DGR), thus enables the production of aggregation-prone and disulfide-containing proteins by refolding from E. coli inclusion bodies, which would not normally be amenable to production in bacteria.

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Details

Published: Jan 19, 2016
Vol/Issue: 6(1)
License: View

Authors

A

Amadeo B. Biter

Department of Investigational Cancer Therapeutics, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA;,

Department of Molecular and Cellular Oncology, University of Texas M.D. Anderson Cancer Center, Houston, TX 77030, USA

Department of Chemistry and Chemical Biology, Howard Hughes Medical Institute, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138

Cite This Article

Amadeo B. Biter, Andres H. de la Peña, Roopa Thapar, et al. (2016). DSF Guided Refolding As A Novel Method Of Protein Production. Scientific Reports, 6(1). https://doi.org/10.1038/srep18906

DSF Guided Refolding As A Novel Method Of Protein Production

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