BioTechniques (Jan 2010)

Purifying natively folded proteins from inclusion bodies using sarkosyl, Triton X-100, and CHAPS

  • Hu Tao,
  • Wenjun Liu,
  • Brandi N. Simmons,
  • Helen K. Harris,
  • Timothy C. Cox,
  • Michael A. Massiah

DOI
https://doi.org/10.2144/000113304
Journal volume & issue
Vol. 48, no. 1
pp. 61 – 64

Abstract

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We describe a rapid, simple, and efficient method for recovering glutathione S-transferase (GST)- and His6-tagged maltose binding protein (MBP) fusion proteins from inclusion bodies. Incubation of inclusion bodies with 10% sarkosyl effectively solubilized >95% of proteins, while high-yield recovery of sarkosyl-solubilized fusion proteins was obtained with a specific ratio of Triton X-100 and CHAPS. We demonstrate for the first time that this combination of three detergents significantly improves binding efficiency of GST and GST fusion proteins to gluthathione (GSH) Sepharose.

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