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Protein engineering to optimize recombinant protein purification.

M Uhlén·T Moks·L Abrahmsén
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Abstract

Genetic approaches have been used to facilitate purification of recombinant proteins, on both a large and a small scale. Based on developments in three different areas: (i) affinity chromatography; (ii) specific cleavage of fusion proteins and (iii) secretion of fusion proteins, a coupled expression/secretion system was designed. It was further improved by protein engineering. Using a synthetic DNA fragment, encoding two IgG-binding domains derived from staphylococcal protein A, gene products were secreted to the culture medium of Escherichia coli and purified with a one-step affinity procedure. The system has been used for large-scale production of biologically active human peptide hormones, to generate peptides for antibody production and to immobilize proteins on solid supports.

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Cloning, MolecularGenesGenetic EngineeringHumansInsulin-Like Growth Factor IInsulin-Like Growth Factor IIPeptide FragmentsRecombinant ProteinsSomatomedinsJournal Article

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M Uhlén, T Moks, L Abrahmsén. 1988. Protein engineering to optimize recombinant protein purification.. https://doi.org/10.1042/bst0160111

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