title: Scaled-up production of recombinant human renin in CHO cells for enzymatic and X-ray structure analysis.
abstract: A process was developed to produce recombinant human renin for X-ray analysis and enzyme inhibition studies. An expression vector containing a human prorenin cDNA and expressing a mouse dihydrofolate reductase selection marker was transfected into dhfr-minus Chinese hamster ovary cells. After selection of cell strains with an increased gene copy number with methotrexate, cultures of the recombinant cells were scaled-up in serum-free media. Major improvements in cellular productivity were achieved by using continuous suspension cultures with cell recycling instead of an adherent culture system or batch-mode suspension cultures. The recombinant zymogen prorenin was purified and preparatively activated with trypsin. Enzymatic properties of the recombinant active renin are described.
authors.affiliation: Biotechnology Department, CIBA-GEIGY Ltd., Basle, Switzerland.
authors.forename: F A
authors.initials: FA
authors.lastname: Asselbergs
authors.forename: J
authors.initials: J
authors.lastname: Rahuel
authors.forename: F
authors.initials: F
authors.lastname: Cumin
authors.forename: C
authors.initials: C
authors.lastname: Leist
mesh_terms: D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003460:Crystallization; D004792:Enzyme Precursors / Q000096:biosynthesis; D006801:Humans; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012083:Renin / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D004792:Enzyme Precursors; D011994:Recombinant Proteins; D012083:Renin