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. 1987;58(1):87-97.
doi: 10.1016/0378-1119(87)90032-1.

A gene fusion system for generating antibodies against short peptides

B Löwenadler  1 , B Jansson, S Paleus, E Holmgren, B Nilsson, T Moks, G Palm, S Josephson, L Philipson, M Uhlén
Affiliations

A gene fusion system for generating antibodies against short peptides

B Löwenadler et al. Gene. 1987.

Abstract

A novel method to obtain specific antibodies against short peptides is described, involving synthesis of the corresponding oligodeoxynucleotides followed by cloning into a new set of fusion vectors, pEZZ8 and pEZZ18, based on two synthetic IgG-binding domains (ZZ) of Staphylococcus aureus protein A. The soluble gene fusion product thus obtained, can be collected from the culture medium of Escherichia coli and rapidly recovered in a one-step procedure by IgG affinity chromatography. The system was used to express a fusion protein consisting of the two Z fragments and the C-terminal part [amino acids (aa) 57-70] of human insulin-like growth factor I (IGF-I). This 16-kDa protein was purified by affinity chromatography on IgG Sepharose and antibodies were raised in rabbits. The fusion protein elicited peptide-specific antibodies, as measured by solid-phase radioimmuno assay and Western blotting, reactive with both synthetic C-terminal peptide and the native human IGF-I protein. The results suggests that the gene fusion system can be used for efficient antibody production against short peptides encoded by synthetic oligodeoxynucleotides.

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