Abstract
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.
MeSH Terms
Escherichia coli/genetics
Gene Expression Regulation
Genetic Engineering
Genetic Vectors
Growth Substances/genetics
Hydrolysis
Insulin/genetics,isolation & purification,metabolism
Insulin Secretion
Mutation
Peptides/genetics,isolation & purification,metabolism
Plasmids
Somatomedins/genetics,isolation & purification,metabolism
Staphylococcal Protein A/genetics
Staphylococcus aureus/genetics
Chemicals
Growth Substances
Insulin
Peptides
Somatomedins
Staphylococcal Protein A
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Nilsson B
Holmgren E
Josephson S
Gatenbeck S
Philipson L
Uhlen M
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