A recombinant human blood coagulation factor ⅷ partially deleted in b region

A technology of human coagulation factor and deletion type, which is applied in the direction of coagulation/fibrinolytic factor, VII factor, blood diseases, etc., and can solve the problems of continuous shortage of FⅧ supply and unsatisfactory rescue medication

Active Publication Date: 2018-06-08
BEIJING NORTHLAND BIOTECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0013] At present, all the plasma-derived FⅧ (plasma-derived FⅧ, pd FⅧ) produced in our country is purified from plasma. However, due to the limitation of raw material plasma supply and plasma purification technology, the annual domestic FⅧ production is only about 60-70 million units. As a result, the supply of domestic FⅧ continues to be tight, and even rescue medication cannot meet the requirements

Method used

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  • A recombinant human blood coagulation factor ⅷ partially deleted in b region
  • A recombinant human blood coagulation factor ⅷ partially deleted in b region
  • A recombinant human blood coagulation factor ⅷ partially deleted in b region

Examples

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Embodiment Construction

[0025] The following examples are only illustrations of the present invention, and do not impose any limitation on the present invention.

[0026] (1) Acquisition and cultivation of production cell lines.

[0027] 1. CHO-DG44 cells.

[0028] Chinese hamster cells - the dihydrofolate reductase (dhfr)-deficient mutant DG44 is similar to the DXB11 mutant cell line used in the early 1980s. The cells require hypoxanthine (or adenine), glycine and thymine for growth, and like all CHO cells, they also require proline for growth. It has the advantage of not containing endogenous dhfr sequence, can be transformed by dhfr gene, and is convenient for screening of positive transformants. For cell lines transformed with the dhfr gene, dhfr-positive transformants are usually screened out with -MEM medium without nucleotides and deoxynucleotides. Since DG44 is a double-deletion mutant that does not contain the hamster dhfr gene, there is no background interference when the cell is introdu...

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Abstract

The invention belongs to the field of biomedicine, in particular to a gene recombinant human coagulation factor VIII for treating hemophilia A. The new gene recombinant human blood coagulation factor VIII product has more stable structure characteristics by performing partial deletion mutation of the B region on the full-length blood coagulation factor VIII.

Description

technical field [0001] The invention belongs to the field of genetic engineering drug production. Through the modification of the full-length coagulation factor Ⅷ (coagulation factor Ⅷ, FⅧ), the recombinant human coagulation factor Ⅷ (recombinanthuman FⅧ, rhFⅧ) with partial deletion of B region was obtained. Experiments have shown that compared with rhFⅧ with complete deletion of B region, the product with partial deletion of B region has a more stable structure and is expected to be developed as a new type of rhFⅧ product for the treatment of hemophilia A. Background technique [0002] Hemophilia (Hemophilia) is a hereditary bleeding disorder in which patients suffer from severe coagulation disorders due to the lack of certain coagulation factors in the blood. The disease is divided into four types: type A, type B, type C and later discovered von Willebrand factor (vonwillibrand factor) lack of pseudohemophilia. Among them, hemophilia A is also called blood coagulation fa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/755A61K38/37A61P7/04
Inventor 许松山聂李亚马素永
Owner BEIJING NORTHLAND BIOTECH
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