Protocol

Protein Folding, Misfolding, and Disease

Volume 752 of the series Methods in Molecular Biology pp 45-57

Date:

Refolding Your Protein with a Little Help from REFOLD

  • Jennifer Phan
  • , Nasrin Yamout
  • , Jason Schmidberger
  • , Stephen P. Bottomley
  • , Ashley M. BuckleAffiliated withDepartment of Biochemistry and Molecular Biology, Monash University Email author 

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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.

Key words

Protein expression Refolding Renaturation Inclusion body Aggregate Refold Misfolding Solubilization