Refolding Your Protein with a Little Help from REFOLD

  • Jennifer Phan
  • Nasrin Yamout
  • Jason Schmidberger
  • Stephen P. Bottomley
  • Ashley M. BuckleEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 752)


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 (, 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 



The authors would like to acknowledge the contribution of all the researchers whose published data has been entered into REFOLD. This work was supported by grants from the National Health and Medical Research Council, the Victorian State Government, and the Victorian Partnership for Advanced Computing. SPB is a Monash University Senior Logan Fellow and NHMRC Senior Research Fellow. AMB is an NHMRC Senior Research Fellow.


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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Jennifer Phan
  • Nasrin Yamout
  • Jason Schmidberger
  • Stephen P. Bottomley
  • Ashley M. Buckle
    • 1
    Email author
  1. 1.Department of Biochemistry and Molecular BiologyMonash UniversityClaytonAustralia

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