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Recombinant CHO cell strain with high expression of human follicle stimulating hormone and construction method thereof

A technology of high expression of human follicle-stimulating hormone, applied in the field of recombinant CHO cell line with high expression of human follicle-stimulating hormone and its construction, which can solve the problems of large differences between individuals and batches, difficulties in industrialization, and low activity.

Pending Publication Date: 2021-10-19
长春生物制品研究所有限责任公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The second type is urine-derived FSH extracted from the urine of postmenopausal women, but there are limited biological resources, large differences between individuals and batches, residual protein components in urine lead to a decrease in the biological activity of FSH, and the production cost is relatively high. high problem
Some scholars have tried to use insect cells and plant cells to express FSH, but there are problems with the glycosylation of natural FSH, the activity is not high, and the industrialization is difficult.

Method used

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  • Recombinant CHO cell strain with high expression of human follicle stimulating hormone and construction method thereof
  • Recombinant CHO cell strain with high expression of human follicle stimulating hormone and construction method thereof
  • Recombinant CHO cell strain with high expression of human follicle stimulating hormone and construction method thereof

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Experimental program
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Effect test

Embodiment

[0021] 1. Artificially synthesized hFSH α subunit and β subunit genes, cloned into pMD-18T vector, ligated and subcloned into pCHO1.0 vector by enzyme digestion, and constructed recombinant co-transfection plasmids pCHO1.0-Fα and pCHO1.0-Fβ and recombinant double expression plasmid pCHO1.0-Fα-Fβ, see figure 1 , figure 2 . The CHO-S suspension cells were transfected, screened by puromycin and MTX pressure, and the expression level was determined by FSH detection kit. High-expression recombinant positive monoclonal cell lines were obtained by multiple rounds of limiting dilution, and the yield of the cell lines was evaluated, and the cell lines with high expression, good activity, and stable expression were selected as engineering cell lines. The cell line construction and screening process is as follows:

[0022] (1) Construction of recombinant expression plasmids

[0023] ①Construction scheme for co-transfection of recombinant plasmids pCHO1.0-Fα and pCHO1.0-Fβ

[0024] ...

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Abstract

The invention relates to the technical field of auxiliary devices of a pregnancy assisting technology, in particular to a recombinant CHO cell strain with high expression of human follicle stimulating hormone and a construction method thereof. The recombinant CHO cell strain is high in purity, low in cost and stable in biological activity, and comprises a CHO-S cell strain, a pCHO1.0 vector, a high expression element and a target protein amino acid sequence.

Description

technical field [0001] The invention relates to the technical field of auxiliary devices for assisted reproductive technology, in particular to a recombinant CHO cell strain highly expressing human follicle-stimulating hormone and a construction method thereof. Background technique [0002] According to WHO statistics, there are about 45 million to 52 million couples affected by infertility in the world. The infertility rate among couples of childbearing age in my country is as high as 10%, involving more than 10 million people. Among them, infertility caused by ovulation disorders Infertility accounts for about 1 / 3, and infertility has become a serious social problem. Human follicle-stimulating hormone is of great significance in the treatment of ovulation disorders, in vitro fertilization and embryo transfer. [0003] FSH is a glycoprotein gonadotropin secreted by basophilic cells of the anterior pituitary gland. It is the core hormone that regulates the growth, sexual matu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C07K14/59C12N15/85
CPCC12N5/0682C07K14/59C12N15/85C12N2510/02
Inventor 曹玉锋常东英张健锋李霄培韩顺子李铮唐剑光时成波
Owner 长春生物制品研究所有限责任公司
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