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.
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[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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