JBM  Vol.4 No.12 , December 2016
The Determination and Evaluation of the Biological Activities for the Commercialization of Recombinant Follicle-Stimulating Hormone in Vitro
Follicle-stimulating hormone (FSH) plays a central role in mammals reproduction, with the actions of FSH mediated by follicle-stimulating hormone receptors (FSHRs) on the surface of target cells. The purposes of this study were to determine and evaluate the biological activities for the commercialization of recombinant follicle-stimulating hormone (rFSH) in vitro through the cellular internalization using cloned 293T-FSHR cell lines as target. Using imaging approaches we have found here that a little fluorescent signal from the surface of the cell transferred to the cytoplasm and accumulated around the nucleus by endocytosis. Compared with the control groups, the commercialization of rFSH have not the significant differences of internalization, but the rFSH have promoted the internalization of the fluorescent, suggested that this detection system might as a protocol for the bioactivity of recombinant therapeutic proteins in vitro.
Cite this paper: Hu, J. , Han, J. , Zhang, X. , Chen, F. , Liu, L. and Zeng, B. (2016) The Determination and Evaluation of the Biological Activities for the Commercialization of Recombinant Follicle-Stimulating Hormone in Vitro. Journal of Biosciences and Medicines, 4, 31-36. doi: 10.4236/jbm.2016.412005.

[1]   Rosenbaum, D.M., Rasmussen, S.G.F. and Kobilka, B.K. (2009) The Structure and Function of G-Protein-Coupled Receptors. Nature, 459, 356-363.

[2]   Lagerström, M.C. and Schiöth, H.B. (2008) Structural Diversity of G Protein-Coupled Receptors and Significance for Drug Discovery. Nature Reviews Drug Discovery, 7, 339-357.

[3]   Radu, A., Pichon, C., Camparo, P., et al. (2010) Expression of Follicle-Stimulating Hormone Receptor in Tumor Blood Vessels. New England Journal of Medicine, 363, 1621-1630.

[4]   Liu, X.M., Chan, H.C., Ding, G.L., et al. (2015) FSH Regulates Fat Accumulation and Redi-stribution in Aging through the Gαi/Ca2+/CREB Pathway. Aging Cell, 14, 409-420.

[5]   Fan, Q.R. and Hendrickson, W.A. (2005) Structure of Human Follicle-Stimulating Hormone in Complex with Its Receptor. Nature, 433, 269-277.

[6]   Moyle, W.R., Lin, W., Myers, R.V., et al. (2005) Models of Glycoprotein Hormone Receptor Interaction. Endocrine, 26, 189-205.

[7]   Perlman, S., van den Hazel, B., Christiansen, J., et al. (2003) Glycosylation of an N-Terminal Extension Prolongs the Half-Life and Increases the in Vivo Activity of Follicle Stimulating Hormone. The Journal of Clinical Endocrinology & Metabolism, 88, 3227-3235.

[8]   Wide, L., Eriksson, K., Sluss, P.M., et al. (2009) Serum Half-Life of Pituitary Gonadotropins Is Decreased by Sulfonation and Increased by Sialylation in Women. The Journal of Clinical Endocrinology & Metabolism, 94, 958-964.

[9]   Verma, R., Boleti, E. and George, A.J.T. (1998) Antibody Engineering: Comparison of Bacterial, Yeast, Insect and Mammalian Expression Systems. Journal of Immunological Methods, 216, 165-181.

[10]   Yin, J., Li, G., Ren, X., et al. (2007) Select What You Need: A Comparative Evalu-ation of the Advantages and Limitations of Frequently Used Expression Systems for Foreign Genes. Journal of Biotechnology, 127, 335-347.

[11]   Dawson, P.E. and Kent, S.B.H. (2000) Synthesis of Native Proteins by Chemical Liga-tion. Annual Review of Biochemistry, 69, 923-960.

[12]   Kent, S.B.H. (2009) Total Chemical Synthesis of Proteins. Chemical Society Re-views, 38, 338-351.

[13]   Aussedat, B., Fasching, B., Johnston, E., et al. (2012) Total Synthesis of the α-Subunit of Human Glycoprotein Hormones: Toward Fully Synthetic Homogeneous Human Follicle- Stimulating Hormone. Journal of the American Chemical Society, 134, 3532-3541.

[14]   Van Damme, M.P., Robertson, D.M., Marana, R., et al. (1979) A Sensitive and Specific in Vitro Bioassay Method for the Measurement of Follicle-Stimulating Hormone Activity. Acta Endocrinologica, 91, 224-237.

[15]   Barrios-De-Tomasi, J., Timossi, C., Merchant, H., et al. (2002) Assessment of the in Vitro and in Vivo Biological Activities of the Human Follicle-Stimulating Isohormones. Molecular and Cellular Endocrinology, 186, 189-198.

[16]   Burgon, P.G., Robertson, D.M., Stanton, P.G., et al. (1993) Immunological Activities of Highly Purified Isoforms of Human FSH Correlate with in Vitro Bioactivities. Journal of Endocrinology, 139, 511-518.

[17]   Albanese, C., Christin-Maitre, S., Sluss, P.M., et al. (1994) Development of a Bioassay for FSH Using a Recombinant Human FSH Receptor and a cAMP Responsive Luciferase Reporter Gene. Molecular and Cellular Endocrinology, 101, 211-219.

[18]   Krishnamurthy, H., Kishi, H., Shi, M., et al. (2003) Postendocytotic Trafficking of the Follicle-Stimulating Hormone (FSH)-FSH Receptor Complex. Molecular Endocrinology, 17, 2162-2176.

[19]   Hunzicker-Dunn, M. and Maizels, E.T. (2006) FSH Signaling Pathways in Immature Granulosa Cells That Regulate Target Gene Expression: Branching out from Protein Kinase A. Cellular Signalling, 18, 1351-1359.

[20]   Wayne, C.M., Fan, H.Y., Cheng, X., et al. (2007) Follicle-Stimulating Hormone Induces Multiple Signaling Cascades: Evidence That Activation of Rous Sarcoma Oncogene, RAS, and the Epidermal Growth Factor Receptor Are Critical for Granulosa Cell Differentiation. Molecular Endocrinology, 21, 1940-1957.

[21]   Casarini, L., Moriondo, V., Marino, M., et al. (2014) FSHR Polymorphism p. N680S Mediates Different Responses to FSH in Vitro. Molecular and Cellular Endocrinology, 393, 83- 91.

[22]   Zhang, Y.L., Guo, K.P., Ji, S.Y., et al. (2016) Development and Characterization of a Novel Long-Acting Recombinant Follicle Stimulating Hormone Agonist by Fusing Fc to an FSH-β Subunit. Human Reproduction, 31, 169-182.