Harnessing Affinity Tags for Recombinant Protein Purification: Concepts, Methods, and Troubleshooting.

Oshadi Edirisinghe, Kaynat Shahzad, Lawrencia Kumi, Zeina Alraawi, Gaëtane Ternier, Patience Salvalina Okoto, Paul Adams, Thallapuranam Krishnaswamy Suresh Kumar

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Article in Methods in molecular biology (Clifton, N.J.), 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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8 authors.

Oshadi EdirisingheCell and Molecular Biology Program, University of Arkansas, Fayetteville, AR, USA.

Kaynat ShahzadDepartment of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR, USA.

Lawrencia KumiCell and Molecular Biology Program, University of Arkansas, Fayetteville, AR, USA.

Zeina AlraawiDepartment of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR, USA.

Gaëtane TernierDepartment of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR, USA.

Patience Salvalina OkotoDepartment of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR, USA.

Paul AdamsCell and Molecular Biology Program, University of Arkansas, Fayetteville, AR, USA.

Thallapuranam Krishnaswamy Suresh KumarCell and Molecular Biology Program, University of Arkansas, Fayetteville, AR, USA. sthalla@uark.edu.

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6 · The paper itself

Abstract

Efficient purification of recombinant proteins is essential for both fundamental biochemical research as well as industrial biotechnology applications. The use of affinity tags has emerged as one of the most powerful and widely adopted method for protein purification. This paper contains recent advancements of design and functions of different affinity tags namely, polyhistidine (His-tag), glutathione S-transferase (GST), small ubiquitin-like modifier (SUMO) tag, maltose-binding protein (MBP), calmodulin-binding protein (CBP), intein-chitin-binding domain tag, N-utilization substance A tag, Halo Tag®, PDZ domain-based tags, thioredoxin tag, polyarginine (Arg-tag) and polyaspartate (Asp-tag) tags, and heparin-binding tag used in recombinant protein purification in bacteria. In addition, we discuss the merits and demerits of each affinity tag.

Indexed as

Chromatography, AffinityRecombinant Fusion ProteinsRecombinant ProteinsEscherichia coliHistidineMaltose-Binding ProteinsHistidineMaltose-Binding ProteinspolyhistidineRecombinant Fusion ProteinsRecombinant ProteinsAffinity chromatographyAffinity tagsBacterial expression systemsFusion proteinsRecombinant protein purificationTag-based purification

Identifiers

PMID42562985

DOI10.1016/j.pep.2010.11.014

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