Highly expressed recombinant human follicle-stimulating hormone from Chinese hamster ovary cells grown in serum-free medium and its effect on induction of folliculogenesis and ovulation

Kim, Dong-Jae; Seok, Seung Hyeok; Baek, Min Won; Lee, Hui Young; Juhn, Jae-Hyeon; Lee, Seungwon; Yun, Minae; Park, Jae‐Hak

doi:10.1016/j.fertnstert.2009.05.009

Cited by 11 publications

(9 citation statements)

References 34 publications

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“…CHO cells do not flourish properly in the absence of FBS, which makes harvesting adequate supplies of recombinant proteins from them difficult. Recently, recombinant gonadotropin production from modified CHO cells has been conducted in the absence of FBS, resulting in proteins that exhibit similar biological activity to previous products produced in the presence of FBS (Kim et al. 2010).…”

Section: Production Methodsmentioning

confidence: 99%

Advances in Recombinant Gonadotropin Production for Use in Bovine Superovulation

Hesser

Morris

Gibbons

2011

Reprod Domestic Animals

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Bovine ovarian hyperstimulation is a process that currently relies on pituitary-derived follicle stimulating hormone (FSH) to facilitate the maturation of multiple follicles to achieve dominance and eventual ovulation. The prevalence of this process, also called superovulation, has more than doubled in the past 10 years, but the efficiency of recovered transferable embryos has remained low at ~6 per collection. The use of pituitary-derived products presents other problems including contamination from other hormones, inconsistencies within and among batches, and the possibility of the spread of disease-transmitting agents. Recombinant gonadotropins have been engineered to yield varieties of FSH and luteinizing hormone from a myriad of heterologous hosts with the resulting products demonstrating various levels of biological activity. Research has also been devoted to alternative delivery methods to reduce the frequency of injections required in current superovulatory protocols. Together, recombinant gonadotropins and alternative delivery approaches potentially provide an economical alternative to the use of pituitary-derived products.

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Section: Production Methodsmentioning

confidence: 99%

Advances in Recombinant Gonadotropin Production for Use in Bovine Superovulation

Hesser

Morris

Gibbons

2011

Reprod Domestic Animals

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“…Since it was not possible to create by restriction-ligation steps the DNA fragment containing the wtEMCV IRES and the FSH chain ORF starting exactly at the start codon 12, the AIB and BIA fragments were constructed from synthetic oligonucleotides (Genescript, Piscataway, NJ, USA). Codon optimization for FSH chains ORFs gave contradictory results earlier [11], the reported expression level from the synthetic ORFs did not exceed that of the full-length natural sequences [12,26], so we used the natural sequences of both ORFs. Two synthetic sequences AIB and BIA were cloned into the p1.1 vector, resulting in the p1.1-FSH-AIB and p1.1-FSH-BIA plasmids (Fig 1A).…”

Section: Resultsmentioning

confidence: 99%

“…The typical FSH production rate reported to date is 0.5–1.5 pg/cell/day [9–11]. Clonal CHO-based cell line, described as being able to produce a “highly expressed” FSH demonstrated a specific productivity of only 0.0085 mIU/cell/48 hours, which corresponds to 0.32 pg/cell/day level [12].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2019

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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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“…Genes encoding the FSH α-subunit and FSH β-subunit, including the signal peptide sequences, were constructed by assembly of synthetic oligonucleotides using the polymerasechain reaction [5]. The primer pairs contained NheI and BamHI enzyme sites, the upstream primer was designed to carry a Nhel restriction site and the downstream primer carried a BamHI restriction site.…”

Section: Construction Of the Rfsh Expression Vectormentioning

confidence: 99%

Antibiotics and ELISA as a Hyphenated Technique for Screening High Production Cell Lines from Transfected HEK293 Cells for the Production of Recombinant Follicle Stimulating Hormone

Hu¹,

Liu²,

Han³

et al. 2017

Proceedings of the 2nd International Conference on Biomedical and Biological Engineering 2017 (BBE 2017)

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Abstract. Recombinant follicle stimulating hormone(rFSH), which play an important role in assisted reproductive technologies. However, the availability of a suitable system for the screening high production cell lines from transfected mammal cells for the production of rFSH is limited. The purpose of this study was to establish a method for screening cloned cell lines with high productivity from transfected HEK293 cells. Here, we adopt the antibiotics in combination with ELISA as a hyphenated technique to obtain production cell lines, and the results indicated that this a simple and efficient method in the cloned cell screening application.

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Highly expressed recombinant human follicle-stimulating hormone from Chinese hamster ovary cells grown in serum-free medium and its effect on induction of folliculogenesis and ovulation

Cited by 11 publications

References 34 publications

Advances in Recombinant Gonadotropin Production for Use in Bovine Superovulation

Advances in Recombinant Gonadotropin Production for Use in Bovine Superovulation

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

Antibiotics and ELISA as a Hyphenated Technique for Screening High Production Cell Lines from Transfected HEK293 Cells for the Production of Recombinant Follicle Stimulating Hormone

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