title: Expression of Bioactive Callithrix jacchus Follicle-Stimulating Hormone in Pichia pastoris.
abstract: Callithrix jacchus (common marmoset) is a New World primate monkey, used as an animal model in biomedical research. Marmoset-specific follicle-stimulating hormone (FSH) preparation is required to improve superovulation protocols and to develop homologous FSH monitoring assays in these monkeys. In this study, we document the large-scale expression of recombinant marmoset FSH in methylotropic yeast, Pichia pastoris. The recombinant preparation was found to be immunologically active in Western blotting and radioimmunoassay. The preparation displayed receptor binding ability in radioreceptor assay. Based on the receptor binding ability, the yield of fermentation was estimated to be 7.2 mg/L. FSH-induced cAMP assay and estradiol assay revealed that the recombinant hormone is able to induce signal transduction. Both immunological and in vitro biological activity of marmoset FSH was found to be comparable to purified human pituitary FSH, which served as reference hormone for these assays. Thus, the study suggests that a Pichia expression system can be used for large-scale expression of bioactive recombinant marmoset FSH.
authors: Division of Structural Biology, National Institute for Research in Reproductive Health (Indian Council of Medical Research), Jehangir Merwanji Street, Parel, Mumbai, 400 012, India.
authors: Susha S
authors: SS
authors: Kutteyil
authors: Bhakti R
authors: BR
authors: Pathak
authors: Rajan R
authors: RR
authors: Dighe
authors: Smita D
authors: SD
authors: Mahale
mesh_terms: D000818:Animals; D002144:Callithrix; D005260:Female; D005640:Follicle Stimulating Hormone / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology*; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D010843:Pichia / Q000235:genetics*; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology
chemical_list: D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone