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

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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.

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Acknowledgments

The research work related to this publication (NIRRH/RA/183/09-2014) was supported by grants received from the Indian Council of Medical Research (BIC/12(10)/2013) and Department of Biotechnology, Government of India (BT/PR4086/AAQ/1/490/2011). Awards of Junior Research Fellowship and Senior Research Fellowship to Ms. Susha S. Kutteyil granted by the Council of Scientific and Industrial Research, India, are greatly acknowledged. The authors thank Dr. J. A. Dias, University at Albany, USA, for providing anti-hFSH β monoclonal antibody. We are also grateful for the help provided by Dr. S. Mukherjee, in-charge of DNA sequencing facility and the technical expertise of Ms. N. Joshi. The technical assistance provided by Mr. B. Kulkarni is also gratefully acknowledged. We also thank Ms. S. Manoharan for the assistance provided with fermentation protocol.

Conflict of Interest

The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.

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Authors and Affiliations

  1. Division of Structural Biology, National Institute for Research in Reproductive Health (Indian Council of Medical Research), Jehangir Merwanji Street, Parel, Mumbai, 400 012, India

    Susha S. Kutteyil, Bhakti R. Pathak & Smita D. Mahale

  2. Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore, 560012, India

    Rajan R. Dighe

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  1. Susha S. Kutteyil
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Correspondence to Smita D. Mahale.

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Kutteyil, S.S., Pathak, B.R., Dighe, R.R. et al. Expression of Bioactive Callithrix jacchus Follicle-Stimulating Hormone in Pichia pastoris . Appl Biochem Biotechnol 176, 399–411 (2015). https://doi.org/10.1007/s12010-015-1583-5

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