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Affinity tags for protein purification

The use of affinity tags for purification of recombinant fusion proteins was first described in 1983 using protein A as a purification tag. The use of affinity tags, including polyhistidine tags (His-tags), has since become widespread as a versatile tool in bioscience, and many tens of thousands of articles have been published on this concept. For the Human Protein Atlas (HPA) effort, the concept was used to generate more than 50,000 recombinant proteins with a histidine affinity tag. These proteins were in the HPA program used for immunizations to generate antibodies to most of the proteins in the human body.

Key publication

  • Nilsson B et al., Immobilization and purification of enzymes with staphylococcal protein A gene fusion vectors. EMBO J. (1985)
    PubMed: 2990908 

Other selected publications

  • Uhlén M et al., Gene fusion vectors based on the gene for staphylococcal protein A. Gene. (1983)
    PubMed: 6313477 DOI: 10.1016/0378-1119(83)90025-2

  • Löwenadler B et al., Production of specific antibodies against protein A fusion proteins. EMBO J. (1986)
    PubMed: 3096719 

  • T. Moks et al., Large-scale affinity purification of human insulin-like growth factor I from culture medium of Escherichia coli. Nat. Biotechnol. (1987)
    DOI: 10.1038/nbt0487-379

  • Moks T et al., Expression of human insulin-like growth factor I in bacteria: use of optimized gene fusion vectors to facilitate protein purification. Biochemistry. (1987)
    PubMed: 3676250 DOI: 10.1021/bi00391a005

  • Ljungquist C et al., Immobilization and affinity purification of recombinant proteins using histidine peptide fusions. Eur J Biochem. (1989)
    PubMed: 2514094 DOI: 10.1111/j.1432-1033.1989.tb15245.x

Figure legend: Basic concepts for using gene fusion to purify a gene product (X) by the use of an affinity tag (A) and an affinity ligand (L). Adapted from Uhlén et al., “Gene fusions for purpose of expression: An introduction”, Methods Enzymol. 185, 129–143 (1990).

Key facts

  • The affinity tag concept was first described in 1983 (protein-A based)
  • The most frequently used affinity tag at present is the histidine-peptide system
  • More than 55,000 gene constructs with affinity tags have been generated in the HPA program
  • A search for “affinity tag” at Google Scholar yields more than 250,000 publications