Production and purification of recombinant human inhibin and activin.
Inhibin and activin are protein hormones with diverse physiological roles including the regulation of pituitary FSH secretion. Like other members of the transforming growth factor-beta gene family, they undergo processing from larger precursor molecules as well as assembly into functional dimers. Isolation of inhibin and activin from natural sources can only produce limited quantities of bioactive protein. To purify large-scale quantities of recombinant human inhibin and activin, we have utilized stably transfected cell lines in self-contained bioreactors to produce protein. These cells produce approximately 200 microg/ml per day total recombinant human inhibin. Conditioned cell media can be purified through column chromatography resulting in dimeric mature 32-34 kDa inhibin A and 28 kDa activin A. The purified recombinant proteins maintain their biological activity as measured by traditional in vitro assays including the regulation of FSH in rat anterior pituitary cultures and the regulation of promoter activity of the activin-responsive promoter p3TP-luc in tissue culture cells. These proteins will be valuable for future analysis of inhibin and activin function and have been distributed to the US National Hormone and Peptide Program.
Pangas
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SA
Department of Neurobiology and Physiology, Northwestern University, Evanston, Illinois 60208, USA.
Woodruff
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TK
D028341:Activins / Q000032:analysis / Q000096:biosynthesis*; D001681:Biological Assay; D019149:Bioreactors; D002460:Cell Line; D002845:Chromatography; D017077:Culture Media, Conditioned / Q000737:chemistry; D006801:Humans; D007265:Inhibins / Q000032:analysis / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012015:Reference Standards
D017077:Culture Media, Conditioned; D011994:Recombinant Proteins; D028341:Activins; D007265:Inhibins