Process for producing a concentrate enriched in coagulation factors VII and VIIa

Claim 1

An improved process for producing a concentrate consisting essentially of blood coagulation factors VII and VIIa and having a specific activity in the range of about 500 to 3000 u/A 280 consisting essentially of the steps of: (a) providing an aqueous solution of plasma proteins containing factors VII and VIIa; (b) contacting the aqueous solution from step (a), adjusted to a pH of about 6 to 8, with an adsorbent having selective affinity for calcium-binding proteins, including factor VII, selected from the group of water-insoluble divalent metal salts, to adsorb calcium-binding proteins; (c) selectively eluting, by techniques selected from step and gradient elution techniques, factors VII and VIIa from the protein-bound adsorbent from step (b) by the addition of a buffer solution containing suitable soluble salts effective to displace the bound proteins and collecting the eluate pool; (d) contacting the eluate pool from step (c), adjusted to a pH of about 3 to 11, with an anionic exchange resin having affinity for the calcium-binding proteins, including factors VII and VIIa, and adsorbing thereon the calcium-binding proteins; and (e) selectively eluting, by techniques selected from step and gradient elution techniques, factors VII and VIIa from the protein-bound adsorbent from step (d) by the addition of a buffer solution containing suitable salts in increasing ionic strength; wherein the improvement comprises using as the starting aqueous solution containing coagulation factors VII and VIIa in step (a) an aqueous wash eluate, containing about 0.1-0.45M of at least one of ammonium bicarbonate, sodium bicarbonate and sodium chloride and containing 0.05-0.2M of citrate ions, said aqueous solution obtained by (i) contacting a Cohn Effluent I plasma fraction containing coagulation factors II, VII, IX and X with an anion exchange resin and (ii) washing the anion exchange resin having adsorbed thereto plasma proteins contained in said plasma fraction to remove those plasma proteins that are less strongly bound to the anion exchange resin so as to be removeable by contacting the anion exchange resin having adsorbed thereto said plasma proteins with a wash solution containing 0.1-0.45M of at least one of ammonium bicarbonate, sodium bicarbonate and sodium chloride.