CHO cell line selection for development of stable cell line producing recombinant human follicle-stimulating hormone
- Authors: Monakhova V.S1, Pigareva N.V1, Simbirtsev A.S1
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Affiliations:
- FGUP «State Research Institute of Highly Pure Biopreparations» of the Federal Biomedical Agency
- Issue: Vol 23, No 3 (2020)
- Pages: 3-7
- Section: Articles
- URL: https://journals.eco-vector.com/1560-9596/article/view/112725
- DOI: https://doi.org/10.29296/25877313-2020-03-01
- ID: 112725
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Abstract
Follicle-stimulating hormone (FSH) is one of the most important hormones of reproductive system and it is widely used as a treatment in an assisted reproductive technology. Choice of productive cell line for recombinant human FSH (rhFSH) protein production is the major issue of biotechnological process. Study objective. The aim of the study was to generate stable cell lines for rhFSH production using different parental CHO cell lines and compare their characteristics. Material and methods. Two CHO suspension parental cell lines were used for stable cell line generation - CHO-K1 and CHO-DXB 11 adapted to suspension conditions. Cell count and viability were analyzed by trypan blue exclusion method. RhFSH concentration was measured by ELISA, protein identity was confirmed by western blot analysis. Results. Four clones generated from CHO-DXB 11 cell line were selected after 3 steps - pool selection, clone selection and methotrexate amplification. Productivity during 6 days of cultivation for the most productive clone CHO-FSH 11/24 was 7,1±0,6 mg/l. Three clones were chosen after transfection and cell cloning of CHO-K1 cells transfected with plasmids containing of FSH α- and β-chains sequences. The highest productivity was detected for CHO-FSH91 clone and it was 10,8±0,9 mg/l. Molecular weight of rhFSH was identical for rhFSH purified from cell culture of optimal clones and commercial preparation Gonal-F (Merck Serono).
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About the authors
V. S Monakhova
FGUP «State Research Institute of Highly Pure Biopreparations» of the Federal Biomedical Agency
Email: v.s.monakhova@hpb.spb.ru
N. V Pigareva
FGUP «State Research Institute of Highly Pure Biopreparations» of the Federal Biomedical Agency
A. S Simbirtsev
FGUP «State Research Institute of Highly Pure Biopreparations» of the Federal Biomedical Agency
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