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Expression of Bioactive Callithrix jacchus Follicle-Stimulating Hormone in Pichia pastoris.
Kutteyil, Susha S; Pathak, Bhakti R; Dighe, Rajan R; Mahale, Smita D.
Afiliação
  • Kutteyil SS; Division of Structural Biology, National Institute for Research in Reproductive Health (Indian Council of Medical Research), Jehangir Merwanji Street, Parel, Mumbai, 400 012, India.
Appl Biochem Biotechnol ; 176(2): 399-411, 2015 May.
Article em En | MEDLINE | ID: mdl-25805018
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.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Pichia / Expressão Gênica / Hormônio Foliculoestimulante Limite: Animals / Female / Humans Idioma: En Revista: Appl Biochem Biotechnol Ano de publicação: 2015 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Pichia / Expressão Gênica / Hormônio Foliculoestimulante Limite: Animals / Female / Humans Idioma: En Revista: Appl Biochem Biotechnol Ano de publicação: 2015 Tipo de documento: Article