Expression of human FSH in Chinese hamster ovarian cells and its activity in a rat model

Authors

Abstract

Background and Objective: FSH is a heterodimeric glycoprotein that performs an important role in the regulation of reproductive processes. Over the past decades, injectable gonadotrophins have played a leading role in the treatment of infertility. The production of FSH in the country is one of the main goals around this drug. The aim of this study was to produce a semi- industrial recombinant FSH.
 
Materials and Methods: In this research study, the FSH gene contained within the pVITRO2-neo-mcs vector was transfected into the cell line of CHO after transformation in DH5α and plasmid purification. The amount of FSH in cell supernatant was measured by ELISA. The FSH protein was purified by HPLC. The presence of FSH protein after purification was confirmed by SDS-PAGE and Western blotting methods. Finally, the clinical effects of purified FSH on the ovary in the rat model were investigated.
 
Results: The expression of the protein was determined by ELISA test at 450 nm that was 43.538 mIU/ml. In the HPLC purification step, the FSH peak was observed at a time interval of about 7.9 minutes, and the purification efficiency was 70%. The existence of a 45 KDa band on the pvdf membrane was confirmed after western blotting of FSH.  Increasing the weight of the ovary after the injection has confirmed the functionality of the protein.
 
Conclusion: At present, the production of recombinant hormone is one of the basic needs of every country. In order to arrive at this aim, a long-term planning is required. We are trying to pave the way for producing human FSH hormone as an effective recombinant drug in the treatment of infertility.
 

Keywords

References

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Daneshvar Medicine
Volume 26, Issue 6 - Serial Number 139
March and April 2019
Pages 1-8

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