School of Biotechnology

  Affinity tag purification
Research groups
Human protein atlas
Mycoplasma mycoides
Antibody proteomics
Alkali stabilization
Tree genomics
Single cell analysis
Biosensor (DNA)
Surface display
Protein folding
In vivo stabilization
Solid phase methods
Protein G
Protein A
Affinity tags

In 1983, the first gene fusion approach for affinity purification of recombinant proteins was described ((1). Fusions to the staphylocoocal protein A (see figure) made it possible to perform one-step purification using IgG-Sepharose (1, 2 and 3). In 1988, a large number of alternative systems were subsequently described (Table 1) all based on the same principle of a one-step purification using a specific affinity tag. The first report on a dual affinity fusion approach was published in 1989 (5). The use of affinity tags for purification of recombinant proteins has since become wide-spread as a versatile tool in bioscience for the affinity capture of recombinant proteins.

Table. The first described gene fusion systems for affinity purifications of recombinant proteins.

Year Affinity tag Ligand for purification Original reference
1983 Protein A IgG Uhlen et al, Gene 23, 369-378
1984 Beta-galactosidase TPEG Ullman, Gene 29, 27-37
1984 Arginine residues Ion-exchange Sassenfeld and Brewer, Bio/Techn. 2: 76-81
1988 Histidine residue Ni2+, Zn2+ Hochuli et al, Bio/Techn. 6: 1321-25
1988 MBP Maltose Di Guan et al, Gene 67: 21-30
1988 GST Glutathion Smith and Johnson, Gene 67:31-40
1988 Protein G (ABD) Albumin Nygren et al, J. Mol. Recogn. 1: 69-74
1988 Flag-peptide Antibody Hopp et al, Bio/Techn. 6: 1204-1210

Key (own) publications:
Uhlen, M., B. Nilsson, B. Guss, M. Lindberg, S. Gatenbeck, and L. Philipson (1983) Gene fusion vectors based on the gene for staphylococcal protein A. Gene 23, 369-378.
Nilsson, B., L. Abrahmsen, and M. Uhlen (1985) Immobilization and purification of enzymes with staphylococcal protein A gene fusion vectors. Embo J 4, 1075-1080.
Lowenadler, B., B. Nilsson, L. Abrahmsen, T. Moks, L. Ljungqvist, E. Holmgren, S. Paleus, S. Josephson, L. Philipson, and M. Uhlen (1986) Production of specific antibodies against protein A fusion proteins. Embo J 5, 2393-2398.
Nygren, P.A., M. Eliasson, L. Abrahmsen, M. Uhlen, and E. Palmcrantz, (1988) Analysis and use of the serum albumin binding domains of streptococcal protein G. J Mol Recognit, 1(2): p. 69-74.
Hammarberg, B., P.A. Nygren, E. Holmgren, A. Elmblad, M. Tally, U. Hellman, T. Moks, and M. Uhlen (1989) Dual affinity fusion approach and its use to express recombinant human insulin-like growth factor II. Proc Natl Acad Sci U S A 86, 4367-4371.
Last updated: 2010-12-22