Protein Affinity Purification using Intein/Chitin Binding Protein Tags

Sarah F. Mitchell, Jon R. Lorsch

Research output: Chapter in Book/Report/Conference proceedingChapter

13 Scopus citations


Isolation of highly purified recombinant protein is essential for a wide range of biochemical and biophysical assays. Affinity purification in which a tag is fused to the desired protein and then specifically bound to an affinity column is a widely used method for obtaining protetin of high purity. Many of these methods have the drawbacks of either leaving the recombinant tag attached to the protein or requiring the addition of a protease which then must be removed by further chromatographic steps. The fusion of a self-cleaving intein sequence followed by a chitin-binding domain (CBD) allows for one-step chromatographic purification of an untagged protein through the thiol-catalyzed cleavage of the intein sequence from the desired protein. The affinity purification is highly specific and can yield pure protein without any undesired N- or C-terminal extensions. This protocol is based on the IMPACT™-System (intein mediated purification with an affinity chitin-binding tag) marketed by New England Biolabs.

Original languageEnglish (US)
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Number of pages15
StatePublished - Jun 20 2015

Publication series

NameMethods in Enzymology
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988


  • CBD
  • Cell harvesting
  • Cell lysis
  • Chitin beads
  • IMPACT™-System
  • Intein/Chitin binding protein tags
  • Protein affinity purification

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology


Dive into the research topics of 'Protein Affinity Purification using Intein/Chitin Binding Protein Tags'. Together they form a unique fingerprint.

Cite this