title: Over-expression of secreted proteins from mammalian cell lines.
abstract: Secreted mammalian proteins require the development of robust protein over-expression systems for crystallographic and biophysical studies of protein function. Due to complex disulfide bonds and distinct glycosylation patterns preventing folding and expression in prokaryotic expression hosts, many secreted proteins necessitate production in more complex eukaryotic expression systems. Here, we elaborate on the methods used to obtain high yields of purified secreted proteins from transiently or stably transfected mammalian cell lines. Among the issues discussed are the selection of appropriate expression vectors, choice of signal sequences for protein secretion, availability of fusion tags for enhancing protein stability and purification, choice of cell line, and the large-scale growth of cells in a variety of formats.
authors: Department of Biochemistry and Molecular Biology, Virginia Commonwealth University, Richmond, Virginia, 23298.
authors: Annamarie C
authors: AC
authors: Dalton
authors: William A
authors: WA
authors: Barton
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007313:Insecta / Q000166:cytology / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D015854:Up-Regulation; D015003:Yeasts / Q000235:genetics
chemical_list: D011994:Recombinant Proteins
keywords: mammalian cell culture; protein expression; receptor; secreted protein