Open Access
PLoS ONE, volume 14, issue 7, pages e0219434
High-level expression of biologically active human follicle stimulating hormone in the Chinese hamster ovary cell line by a pair of tricistronic and monocistronic vectors
Kovnir Sergey V
1
,
Khodak Yulia A
1
,
Polzikov Mikhail A.
2
,
Nikitina Victoria
3
,
Skryabin Konstantin G.
1
,
2
IVFarma LLC, Moscow, Russia
|
Publication type: Journal Article
Publication date: 2019-07-05
PubMed ID:
31276557
Multidisciplinary
Abstract
Recombinant human follicle stimulating hormone (FSH), produced in Chinese hamster ovary (CHO) cells, is widely used for treatment of fertility disorders and is subject to biosimilars development. Cell lines with high specific productivities may simplify the FSH production process. Here, we used our previously established expression system based on vector p1.1 to create new cell lines secreting heterodimeric FSH protein. To this end, we linked open reading frames of both FSH subunits by the wild-type internal ribosome entry site from the encephalomyocarditis virus (EMCV IRES). Intact and double-negative for the dihydrofolate reductase CHO cells were stably transfected by the FSH-coding plasmids. Stably transfected intact cells showed higher level of the FSH secretion and were utilized for subsequent methotrexate-driven transgene amplification, which doubled their productivity. The excess of the free α-subunit was corrected by transfecting the cells by the additional p1.1-based plasmid encoding the β-subunit of the FSH. Clonal cell lines obtained secreted mostly the heterodimeric FSH and possessed specific productivities up to 12.3±1.7 pg/cell/day. Candidate clonal cell line C-P1.3-FSH-G4 maintained a constant specific productivity for at least 2 months of culturing without the section pressure. The resulting FSH protein conformed to the international pharmaceutical quality criteria as evidenced by the receptor binding kinetics, distribution pattern of hormone isoforms and biological activity. In conclusion, our expression system offers a simple and cost-effective approach to production of FSH.
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Citations by publishers
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- We do not take into account publications that without a DOI.
- Statistics recalculated only for publications connected to researchers, organizations and labs registered on the platform.
- Statistics recalculated weekly.
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Orlova N. A. et al. High-level expression of biologically active human follicle stimulating hormone in the Chinese hamster ovary cell line by a pair of tricistronic and monocistronic vectors // PLoS ONE. 2019. Vol. 14. No. 7. p. e0219434.
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Orlova N. A., Kovnir S. V., Khodak Y. A., Polzikov M. A., Nikitina V., Skryabin K. G., Vorobiev I. I. High-level expression of biologically active human follicle stimulating hormone in the Chinese hamster ovary cell line by a pair of tricistronic and monocistronic vectors // PLoS ONE. 2019. Vol. 14. No. 7. p. e0219434.
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TY - JOUR
DO - 10.1371/journal.pone.0219434
UR - https://doi.org/10.1371%2Fjournal.pone.0219434
TI - High-level expression of biologically active human follicle stimulating hormone in the Chinese hamster ovary cell line by a pair of tricistronic and monocistronic vectors
T2 - PLoS ONE
AU - Orlova, Nadezhda A.
AU - Kovnir, Sergey V
AU - Khodak, Yulia A
AU - Polzikov, Mikhail A.
AU - Nikitina, Victoria
AU - Skryabin, Konstantin G.
AU - Vorobiev, Ivan I
PY - 2019
DA - 2019/07/05 00:00:00
PB - Public Library of Science (PLoS)
SP - e0219434
IS - 7
VL - 14
PMID - 31276557
SN - 1932-6203
ER -
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@article{2019_Orlova,
author = {Nadezhda A. Orlova and Sergey V Kovnir and Yulia A Khodak and Mikhail A. Polzikov and Victoria Nikitina and Konstantin G. Skryabin and Ivan I Vorobiev},
title = {High-level expression of biologically active human follicle stimulating hormone in the Chinese hamster ovary cell line by a pair of tricistronic and monocistronic vectors},
journal = {PLoS ONE},
year = {2019},
volume = {14},
publisher = {Public Library of Science (PLoS)},
month = {jul},
url = {https://doi.org/10.1371%2Fjournal.pone.0219434},
number = {7},
pages = {e0219434},
doi = {10.1371/journal.pone.0219434}
}
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MLA
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Orlova, Nadezhda A., et al. “High-level expression of biologically active human follicle stimulating hormone in the Chinese hamster ovary cell line by a pair of tricistronic and monocistronic vectors.” PLoS ONE, vol. 14, no. 7, Jul. 2019, p. e0219434. https://doi.org/10.1371%2Fjournal.pone.0219434.