RECOMBINANT PLASMID DNA pAP227 CODING POLYPEPTIDE OF RECOMBINANT FACTOR VIII OF HUMAN BLOOD COAGULABILITY, LINE OF CELLS Cricetulus griseus CHO, 2H5-PRODUCER OF RECOMBINANT FACTOR VIII OF HUMAN BLOOD COAGULABILITY, AND METHOD FOR OBTAINING POLYPEPTIDE HAVING ACTIVITY OF RECOMBINANT FACTOR VIII Russian patent published in 2013 - IPC C12N15/11 C12N15/63 C12N5/10 C12P21/02 C07K1/16 

Abstract RU 2500818 C1

FIELD: biotechnologies.

SUBSTANCE: invention can be used for obtaining recombinant human blood coagulability factor VIII with deletion of B-domain (hFVIII-BDD). Recombinant plasmid DNA pAP227 coding polypeptide with sequence hFVIII-BDD also including MAR - binding area to nuclear matrix of lysozyme gene of birds, virus transmission enhancer CMV, internal translation initiation site IRES of encephalomyocarditis virus, gene DHFR of a mouse, a polyadenylation signal of virus SV40, gene of aminoglycoside-3'-phosphotransferase providing stability to geneticin (Neo) and a cassette for expression in bacteria cells of gene of β-lactamase providing stability to ampicillin, cells of line Cricetulus griseus CHO DHFR(-) are obtained so that there produced is cell line Cricetulus griseus CHO 2H5 producing recombinant hFVIII-BDD with highly stable yield at the level of about 20 IU/ml/24 h. Cultivation of cells-producers is performed in medium DME/F12 containing 2-4% of Fetal Bovine Serum, 1% of dimethylsulphoxide and 50 IU/l of insulin.

EFFECT: improvement of the method.

4 cl, 5 dwg, 9 ex

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RU 2 500 818 C1

Authors

Grigor'Eva Ol'Ga Vasil'Evna

Zaval'Nyj Mikhail Aleksandrovich

Karpov Andrej Pavlovich

Petrov Andrej Vladimirovich

Fabrichnyj Igor' Pavlovich

Shuster Aleksandr Mikhajlovich

Dates

2013-12-10Published

2012-07-19Filed