title: Recombinant human transforming growth factor-beta 1: expression by Chinese hamster ovary cells, isolation, and characterization.
abstract: A partial cDNA clone encoding the gene for human transforming growth factor-beta 1 (TGF-beta 1) was isolated from a human bladder carcinoma cell line (5637) cDNA library. Following restriction enzyme processing and ligation of synthetic oligonucleotide linkers, the gene was inserted into a plasmid and transfected into Chinese hamster ovary cells. Clonal selection and growth conditions resulted in a method for production of recombinant human TGF-beta 1 at 7 mg/liter in conditioned cell medium. Through a combination of low pH treatment, cation-exchange chromatography, and salt precipitation, the recombinant human TGF-beta 1 was purified in milligram amounts to > 95% purity in a yield of about 36%. Purification to homogeneity was accomplished by chromatography on C18 silica gel. Amino acid analysis, N-terminal sequencing, and growth inhibition assays indicate identity with the molecule from human platelets.
authors: Amgen, Inc., Amgen Center, Thousand Oaks, California 91320.
authors: L
authors: L
authors: Bourdrel
authors: C H
authors: CH
authors: Lin
authors: S L
authors: SL
authors: Lauren
authors: R H
authors: RH
authors: Elmore
authors: B J
authors: BJ
authors: Sugarman
authors: S
authors: S
authors: Hu
authors: K R
authors: KR
authors: Westcott
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001483:Base Sequence; D001681:Biological Assay / Q000379:methods; D016466:CHO Cells; D002455:Cell Division / Q000187:drug effects; D002461:Cell Line, Transformed; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D006224:Cricetinae; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D017421:Sequence Analysis; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D000596:Amino Acids; D003470:Culture Media; D016212:Transforming Growth Factor beta