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Method for producing recombinant human coagulation factor VII by using rabbit mammary gland reactor platform

A technology of mammary gland reactor and coagulation factor, which is applied in the field of production of recombinant human coagulation factor VII, can solve the problems of high production cost and low protein expression level, and achieve the effect of low cost, short cultivation cycle and good activity

Active Publication Date: 2014-01-01
LANNUO BIOTECH WUXI
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Problems solved by technology

The recombinant protein products of coagulation factor VII currently on the market use the hamster larval kidney cell (CHO) culture system for protein expression, and the protein expression level is low, resulting in extremely high production costs

Method used

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  • Method for producing recombinant human coagulation factor VII by using rabbit mammary gland reactor platform
  • Method for producing recombinant human coagulation factor VII by using rabbit mammary gland reactor platform
  • Method for producing recombinant human coagulation factor VII by using rabbit mammary gland reactor platform

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Experimental program
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Embodiment 1

[0033] (1) Preparation of the seventh blood coagulation factor mammary gland expression vector:

[0034] a. Seventh blood coagulation factor rhFⅦ cDNA amplification: the recombinant seventh blood coagulation factor rhFⅦ cDNA structure is as follows figure 1 As shown, including light chain (light chain) and heavy chain (heavy chain), the end of the light chain is externally connected with a signal peptide S, and the signal peptide S includes the original secretion signal peptide (19 amino acid polypeptide) of the seventh coagulation factor hFVII With β-casein signal peptide (19 amino acid polypeptides), at the same time, rhFⅦ cDNA outside bGH polyA (not shown in the figure), bGH polyA can increase the protein translation level and optimize the expression effect. The hFVII fragment was amplified from the human cDNA library, and two sets of primers were prepared, one set was pBC1hFVII-f / pBC1hFVII-r, and the other set was pBC1hFVII-f2 / pBC1hFVII-r. figure 1 The arrows shown i...

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Abstract

The invention provides a method for producing a recombinant human coagulation factor VII by using a rabbit mammary gland reactor platform. CDNA (Complementary deoxyribonucleic acid) of the recombinant human coagulation factor VII is guided or integrated to a chromosome gene of a rabbit, and the obtained transgenic rabbit expresses recombinant human coagulation factor VII protein in a mammary gland reactor. The transgenic rabbit has relatively high recombinant human coagulation factor VII protein expression level in the mammary gland reactor, the protein activity is good, and the production cost is reduced.

Description

technical field [0001] The invention belongs to the field of preparation of recombinant protein medicine by non-human mammals, and specifically relates to a method for producing recombinant human seventh blood coagulation factor using a rabbit mammary gland reactor platform. Background technique [0002] Hemophilia is the most common bleeding disorder in humans. It is lifelong, recurrent and fatal. About one in every 5000-10000 men is a hemophiliac. Hemophilia is caused by the absence of certain functional coagulation factors in the blood, and the most effective treatment for hemophilia is direct injection of coagulation factors. Now the most common blood coagulation factors used for treatment are coagulation factors VIII and IX. However, after frequent treatment with exogenous coagulation factors VIII and IX, the patient develops resistance to these coagulation factors, or neutralizes them. Sexual antibodies, thereby increasing the risk of coagulation factor therapy, leadi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85
Inventor 杜福良杨澜薛非安礼友
Owner LANNUO BIOTECH WUXI
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