Abstract
At present the protein expression systems used commonly by researchers incorporate an affinity tail fused to the protein of interest. These affinity tails provide a convenient and efficient method for the purification of the expressed fusion protein using affinity chromatography. Many different affinity tails have been developed and a few of the commonly used fusion protein expression systems are listed in Table 1. The plasmid expression vectors for the production of fusion proteins in various hosts are available commercially and with the advent of the polymerase chain reaction (PCR) any gene of known sequence can be cloned into any expression vector. Most affinity tails are linked to the N-terminus of the protein of interest, but some affinity tails are able to be linked to either the N- or C-terminus of the protein of interest. The choice of affinity tail to use for the expression of any particular protein is empirical since the factors leading to the high expression of recombinant proteins in foreign hosts have yet to be elucidated.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Chang J.-Y. (1985) Thrombin specificity. Requirement for apolar amino acids adjacent to the thrombin cleavage site of polypeptide substrate Eur J Biochem. 151, 217–224.
Walker P. A., Leong L. E., Ng P. W, Tan S. H, Waller S, Murphy D., and Porter A. G (1994) Efficient and rapid purification of proteins using recombinant fusion proteases Bio/Technology 12, 601–605
Urzainqui A. and Carrasco L (1989) Degradation of cellular proteins during poliovirus infectron studies by two-dimensional gel electrophoresis J Virol 63, 4729–4735
Cordingley M. G., Callahan P L., Sardana V. V, Garsky V. M, and Colonno R. J. (1990) Substrate requirements of human rhinovirus 3C protease for peptide cleavage in vitro J Biol Chem 265, 9062–9065
Smith D. B and Johnson K. S (1988) Single step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67, 31–40
Smith M. C., Furman T. C., Ingolia T. D., and Pidgeon C. (1988) Chelating peptide-immobilized metal ion affinity chromatography J Biol Chem 263, 7211–7215.
Van Dyke M. W., Sirito M., and Sawadogo M (1992) Single step purification of bacterially expressed polypeptides containing an oligo-histidine domain. Gene 111, 99–104.
di Guan C., Li P, Riggs P. D., and Inouye H. (1988) Vectors that facilitate the expression and purification of foreign peptides in Escherichia coli by fusion to maltose-binding protein Gene 67, 21–30
Lowenadler B., Jansson B., Paleus S, Holingren E., Nilsson B., Moks T., Palm G, Josephson S, Philipson L., and Uhlen M (1987) A gene fusion system for generating antibodies against short peptides Gene 58, 87–97
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1998 Humana Press Inc.
About this protocol
Cite this protocol
Leong, L.E.C. (1998). The Use of Recombinant Fusion Proteases in the Affinity Purification of Recombinant Proteins. In: Reischl, U. (eds) Molecular Diagnosis of Infectious Diseases. Methods in Molecular Medicine™, vol 13. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-485-2:323
Download citation
DOI: https://doi.org/10.1385/0-89603-485-2:323
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-0-89603-485-3
Online ISBN: 978-1-59259-597-6
eBook Packages: Springer Protocols