DSF Guided Refolding As A Novel Method Of Protein Production

Scientific Reports
Amadeo B BiterKevin J Phillips

Abstract

The 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.

References

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Citations

May 25, 2016·Scientific Reports·Lu ZhaiBin Xu
Sep 25, 2019·Analytical Biochemistry·Amadeo B BiterMaria Elena Bottazzi
Jan 20, 2020·Trends in Biotechnology·Priyank SinghviAmulya K Panda
Feb 6, 2018·ACS Omega·William T BoothMaksymilian Chruszcz

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Methods Mentioned

BETA
protein folding
NMR

Software Mentioned

Felix
DGR

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