Refolding your protein with a little help from REFOLD

Methods Mol Biol. 2011:752:45-57. doi: 10.1007/978-1-60327-223-0_4.

Abstract

The expression and harvesting of proteins from insoluble inclusion bodies by solubilization and refolding is a technique commonly used in the production of recombinant proteins. Despite the importance of refolding, publications in the literature are essentially ad hoc reports consisting of a dazzling array of experimental protocols and a diverse collection of buffer cocktails. For the protein scientists, using this information to refold their protein of interest presents enormous challenges. Here, we describe some of the practical considerations in refolding and present several standard protocols. Further, we describe how refolding procedures can be designed and modified using the information in the REFOLD database (http://refold.med.monash.edu.au), a freely available, open repository for protocols describing the refolding and purification of recombinant proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Buffers
  • Chelating Agents / chemistry
  • Chromatography, Gel
  • Databases, Protein*
  • Dialysis
  • Disulfides / chemistry
  • Inclusion Bodies / chemistry
  • Inclusion Bodies / genetics
  • Nickel / chemistry
  • Protein Engineering
  • Protein Refolding*
  • Recombinant Proteins / chemistry*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Sequence Homology, Amino Acid
  • Solubility

Substances

  • Buffers
  • Chelating Agents
  • Disulfides
  • Recombinant Proteins
  • Nickel