Simple and Efficient Purification of Recombinant Proteins Using the Heparin-Binding Affinity Tag.
Curr Protoc Protein Sci
; 90: 6.16.1-6.16.13, 2017 11 01.
Article
de En
| MEDLINE
| ID: mdl-29091276
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.
Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Agarose
/
Protéines de fusion recombinantes
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Protéines du sang
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Protéines de transport
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Chromatographie d'affinité
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Peptides antimicrobiens cationiques
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Escherichia coli
Langue:
En
Journal:
Curr Protoc Protein Sci
Année:
2017
Type de document:
Article