Bio/Technology
5, 379 - 382 (1987)
doi:10.1038/nbt0487-379
Large−Scale Affinity Purification of Human Insulin−Like Growth Factor I from Culture Medium of Escherichia ColiTomas Moks1, Lars Abrahmsén1, Björn Österlöf2, Staffan Josephson2, Martin Östling3, Sven-Olof Enfors1, Ingalill Persson4, Björn Nilsson1
& Mathias Uhlén1
1Department of Biochemistry and Biotechnology, The Royal Institute of Technology, S-100 44 Stockholm, Sweden.
2Kabigen AB, S-112 87 Stockholm, Sweden.
3Pharmacia Biotechnology, S-751 82 Uppsala, Sweden.
4Alfa Laval AB, Biotech Center, S-147 00 Tumba, Sweden. A genetic approach to facilitate large−scale production and down stream processing of heterologous proteins expressed in bacteria is described. The gene is fused after a synthetic fragment encoding two IgG−binding domains derived from staphylococcal protein A. The fusion product is secreted to the growth medium of Escherichia coli, and purified using the IgG−binding moiety as affinity "tail". We demonstrate that the expression system can be used for the production of biologically active human insulin−like growth factor I in a 1000 litre scale. The process includes fermentation, cell separation, affinity chromatography on IgG Sepharose Fast Flow and site specific chemical cleavage of the fusion protein.
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