Expression of Bioactive Callithrix jacchus Follicle-Stimulating Hormone in Pichia pastoris

Kutteyil, Susha S and Pathak, Bhakti R and Dighe, Rajan R and Mahale, Smita D (2015) Expression of Bioactive Callithrix jacchus Follicle-Stimulating Hormone in Pichia pastoris. In: APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY, 176 (2). pp. 399-411.

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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.

Item Type:	Journal Article
Publication:	APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
Publisher:	HUMANA PRESS INC
Additional Information:	Copy right for this article belongs to the HUMANA PRESS INC, 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA
Keywords:	Callithrix jacchus; Follicle-stimulating hormone; Pichia pastoris; Cloning; Expression; Fermentation; Bioactivity
Department/Centre:	Division of Biological Sciences > Molecular Reproduction, Development & Genetics
Date Deposited:	26 Jun 2015 06:50
Last Modified:	26 Jun 2015 06:50
URI:	http://eprints.iisc.ac.in/id/eprint/51777

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