Zone B partially-deleted type recombinant human blood coagulation factor VIII

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

Active Publication Date: 2015-11-04
BEIJING NORTHLAND BIOTECH
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  • Abstract
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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 plasma-derived FⅧ, but due to the limitation of raw material plasma supply and plasma purification

Method used

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  • Zone B partially-deleted type recombinant human blood coagulation factor VIII
  • Zone B partially-deleted type recombinant human blood coagulation factor VIII
  • Zone B partially-deleted type recombinant human blood coagulation factor VIII

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 biological medicines, and in particular relates to a gene recombinant human blood coagulation factor VIII for treating hemophilia A. By performing zone B partially-deleted type mutation on a full-length blood coagulation factor VIII, the obtained novel gene recombinant human blood coagulation factor VIII product has a characteristic that the structure is more stable.

Description

technical field [0001] The invention belongs to the field of genetic engineering drug production. Through the transformation of the full-length coagulation factor VIII (coagulation factor VIII, FVIII), the recombinant human coagulation factor VIII (recombinant human 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...

Claims

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Application Information

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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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