Unit

UNIT 26.4 Recombinant Protein Purification by Self-Cleaving Elastin-like Polypeptide Fusion Tag

  1. Wan-Yi Wu,
  2. Baley A. Fong,
  3. Allison G. Gilles,
  4. David W. Wood

Published Online: 1 NOV 2009

DOI: 10.1002/0471140864.ps2604s58

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Wu, W.-Y., Fong, B. A., Gilles, A. G. and Wood, D. W. 2009. Recombinant Protein Purification by Self-Cleaving Elastin-like Polypeptide Fusion Tag. Current Protocols in Protein Science. 58:26.4:26.4.1–26.4.18.

Author Information

  1. Princeton University, Princeton, New Jersey

Publication History

  1. Published Online: 1 NOV 2009
  2. Published Print: NOV 2009

Abstract

This unit presents a rapid and simple method for the nonchromatographic purification of recombinant proteins expressed in E. coli. This method relies on a thermally responsive elastin-like polypeptide (ELP) tag, where the tagged protein is precipitated using a mild temperature shift. The tag is then induced to self-cleave by a mild pH shift and is subsequently removed by a final thermal precipitation. The result is a purified native protein target, without the requirement for affinity apparatus or protease removal of the tag. This protocol describes the required cloning methods to insert a given target into the expression vector, as well as the general method for purifying the resulting expressed protein. Curr. Protoc. Protein Sci. 58:26.4.1-26.4.18. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • intein;
  • elastin-like polypeptide;
  • protein purification;
  • Gateway cloning;
  • nonchromatographic bioseparations