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ACS Comb Sci. 2013 Aug 12;15(8):387-92. doi: 10.1021/co400013v. Epub 2013 Jun 10.

High-throughput thermal scanning for protein stability: making a good technique more robust.

Author information

1
Collaborative Crystallisation Centre (C3), Division of Materials Science and Engineering, Commonwealth Scientific and Industrial Research Organisation (CSIRO) , Parkville, Victoria, Australia.

Abstract

We present a high-throughput approach to help define experimental formulations that enhance protein stability, which is based on differential scanning fluorimetry (DSF). The method involves defining the thermal stability of a protein against a screen of 13 buffer systems, systematically sampling pH from 5.0 to 9.0 at high and low salt concentrations, using both redundancy and extensive controls to make the method robust. The screen allows rapid determination of a suitable base formulation for protein samples, and is particularly useful for difficult samples: those that are rapidly degraded or cannot be sufficiently concentrated for downstream analyses. Data obtained from three samples in this assay illustrate the vastly different values for thermal stability that can be obtained from different formulations. This approach is simple to interpret and reliable enough that it has been implemented as a service through the Collaborative Crystallisation Centre (C3).

PMID:
23710551
DOI:
10.1021/co400013v
[Indexed for MEDLINE]

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