Simple and Efficient Purification of Recombinant Proteins Using the Heparin-Binding Affinity Tag
by Jayanthi, Srinivas; Gundampati, Ravi Kumar; Kumar, Thallapuranam Krishnaswamy Suresh
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 mol. 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 chromatog. using a simple sodium chloride gradient to elute the bound fusion protein. In addition, owing to the high d. of pos. 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 chem. denaturants, including urea. Addnl., polyclonal antibodies raised against the affinity tag can be used to detect HBP-fused target proteins with high sensitivity.
- Journal
- Current Protocols in Protein Science
- Volume
- 90
- Issue
- 1
- Year
- 2017
- URL
- https://dx.doi.org/10.1002/cpps.41
- ISBN/ISSN
- 1934-3663; 1934-3655
- DOI
- 10.1002/cpps.41