Simple and Efficient Purification of Recombinant Proteins Using the Heparin‐Binding Affinity Tag. (13th February 2018)

• Record Type: Journal Article
• Authors: Jayanthi, Srinivas; Gundampati, Ravi Kumar; Kumar, Thallapuranam Krishnaswamy Suresh
• Editors: Coligan, John E.; Dunn, Ben M.; Speicher, David W.; Wingfield, Paul T.
• Is Part Of: Current protocols in protein science. Volume 90(2017)
• Page Start: 6.16.1
• Page End: 6.16.13
• Publication Date: 2018-02-13
• Subjects: affinity chromatography -- fusion tag -- heparin sepharose -- recombinant protein -- resin; Proteins -- Laboratory manuals; Proteins; Clinical Laboratory Techniques; Genetic Techniques; Immunologic Techniques; Proteins; Laboratory manuals; 572.6028
• DOI: 10.1002/cpps.41
• Languages: English
• ISSNs: 1934-3655

Abstract

Abstract: Heparin, a member of the glycosaminoglycan family, is known to interact with more than 400 different types of proteins. For the past few decades, significant progress has been made to understand the molecular details involved in heparin‐protein interactions. Based on the structural knowledge available from the FGF1‐heparin interaction studies, we have designed a novel heparin‐binding peptide (HBP) affinity tag that can be used for the simple, efficient, and cost‐effective purification of recombinant proteins of interest. HBP‐tagged fusion proteins can be purified by heparin Sepharose affinity chromatography using a simple sodium chloride gradient to elute the bound fusion protein. In addition, owing to the high density of positive charges on the HBP tag, recombinant target proteins are preferably expressed in their soluble forms. The purification of HBP‐fusion proteins can also be achieved in the presence of chemical denaturants, including urea. Additionally, polyclonal antibodies raised against the affinity tag can be used to detect HBP‐fused target proteins with high sensitivity. © 2017 by John Wiley & Sons, Inc.