Protein engineering to optimize recombinant protein purification.
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.
Uhlén
M
M
Department of Biochemistry, Royal Institute of Technology, Stockholm, Sweden.
Moks
T
T
Abrahmsén
L
L
D003001:Cloning, Molecular / Q000379:methods*; D005796:Genes; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000235:genetics* / Q000302:isolation & purification; D007335:Insulin-Like Growth Factor II / Q000235:genetics* / Q000302:isolation & purification; D010446:Peptide Fragments / Q000032:analysis; D011994:Recombinant Proteins / Q000302:isolation & purification*; D013002:Somatomedins / Q000235:genetics*
D010446:Peptide Fragments; D011994:Recombinant Proteins; D013002:Somatomedins; D007334:Insulin-Like Growth Factor I; D007335:Insulin-Like Growth Factor II