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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: |
| 1. |
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. |
| 2. |
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. |
| 3. |
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. |
| 4. |
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. |
| 5. |
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. |
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