Open AccessEfficient secretion and purification of human insulin-like growth factor I with a gene fusion vector inStaphylococci
Author(s) -
Björn Nilsson,
Erik Holmgren,
Staffan Josephson,
Sten Gatenbeck,
Lennart Philipson,
Mathias Uhlén
Publication year - 1985
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/13.4.1151
Subject(s) - biology , fusion protein , cleavage (geology) , biochemistry , secretion , affinity chromatography , protein a , fusion gene , expression vector , gene , microbiology and biotechnology , recombinant dna , antibody , genetics , paleontology , fracture (geology) , enzyme
A novel approach for production of small polypeptides, using a staphylococcal protein A vector, is described. This system is used to express, secrete and purify human insulin-like growth factor I (IGF-I). A fusion protein consisting of protein A and IGF-I is recovered in high yield by passing the culture medium through an IgG affinity column. Using site-specific mutagenesis an acid labile asp-pro cleavage site was introduced at the fusion point between the two proteins. The protein A "tail" can thereby be removed from the affinity purified fusion protein by chemical cleavage releasing biologically active IGF-I molecules.