Factor IX preparations uncontaminated by plasma components or po

Details Factor IX preparations uncontaminated by plasma components or po Factor IX preparations uncontaminated by plasma components or po

1991-09-23

1992-12-15

Wax, Robert A.

Drug, bio-affecting and body treating compositions

Enzyme or coenzyme containing

Hydrolases

435219, 514 8, 530381, 530384, C07K 1300, A61K 3754, A61K 3748, C12N 948

Patent

active

051715694

ABSTRACT:
The blood-clotting protein, factor IX, is synthesized in the bod in liver cells, where it undergoes three distinct types of post-translational modification before it is secreted into the bloodstream as a 415 amino acid long protein. It is therefore a difficult protein to produce by recombinant DNA technology in a highly biologically active form. Nevertheless, such a result has been achieved by the present invention in which typically factor IX cDNA in a plasmid is linearized and inserted into an expression vector having a promoter sequence of SV40 early gene, an SV40 polyadenylation sequence, the TK/NEO selectable marker and an ampicillin resistance gene. Mammalian cells such as from a dog kidney or rat liver are transfected by the calcium phosphate precipitation method. High levels of factor IX in a fully or near-fully biologically active form, useful as a plasma-free preparation for treatment of patients suffering from Christmas Disease (haemophilia B), are obtainable without recourse to poxvirus vectors which would contaminate the protein.

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