Virus Filtration of Liquid Factor VII Compositions

Abstract

The present invention relates to a novel method for improving the viral safety of liquid Factor VII compositions, in particular those comprising active Factor VII polypeptides (a Factor VIIa polypeptide).

1 . A method for removing viruses from a liquid Factor VII composition comprising one or more Factor VII polypeptides wherein at least 5% of the Factor VII polypeptides are in an activated form, the method comprising subjecting the solution to nanofiltration using a nanofilter having a pore size of 80 nm or less. 
     
     
         2 . The method according to  claim 1 , wherein at least about 50% of the mass of the Factor VII polypeptides are in activated form. 
     
     
         3 . The method according to  claim 1 , wherein the liquid composition has a pH of in the range of 7.0-9.5. 
     
     
         4 . The method according to  claim 3 , wherein the pH is in the range of 8.3-8.7. 
     
     
         5 . The method according to  claim 1 , wherein the concentration of the Factor VII polypeptide(s) in the liquid composition is in the range of 0.01-5 mg/mL. 
     
     
         6 . The method according to  claim 1 , wherein the pore size of the nanofilter is 50 nm or less. 
     
     
         7 . The method according to  claim 1 , wherein the membrane of the nanofilter is manufactured from one or more materials selected from the group consisting of: cuprammonium regenerated cellulose, hydrophilic polyvinylidene fluoride (PVDF), composite PVDF, surface modified PVDF, and polyether sulfone. 
     
     
         8 . The method according to  claim 1 , wherein the pore size of the nanofilter is at the most 50 nm, e.g. at the most 30 nm, such as in the range of 10-30 nm. 
     
     
         9 . The method according to  claim 1 , wherein the liquid Factor VII composition is obtained from a cell culture supernatant. 
     
     
         10 . The method according to  claim 1 , wherein the liquid composition is substantially serum-free. 
     
     
         11 . The method according to  claim 1 , wherein the Factor VII polypeptide(s) is/are produced by cell culture in the presence of bovine or foetal calf serum. 
     
     
         12 . A method for removing viruses from a liquid Factor VII composition comprising one or more Factor VII polypeptides, the liquid composition being substantially serum-free, the method comprising subjecting said solution to nanofiltration using a nanofilter having a pore size of at the most 80 nm. 
     
     
         13 . The method according to  claim 12 , wherein the Factor VII polypeptide(s) is/are produced by cell culture in CHO cells in a medium free from any components of animal origin. 
     
     
         14 . The method according to  claim 12 , wherein at least 5% of the one or more Factor VII polypeptides are in the activated form. 
     
     
         15 . The method according to  claim 12 , wherein the liquid composition has a pH of in the range of 8.3-8.7. 
     
     
         16 . The method according to  claim 12 , wherein the concentration of the Factor VII polypeptide(s) in the liquid composition is in the range of 0.05-2.0 mg/mL. 
     
     
         17 . The method according to  claim 12 , wherein the pore size of the nanofilter is 30 nm or less. 
     
     
         18 . The method according to  claim 12 , wherein the membrane of the nanofilter is manufactured from one or more materials selected from the group consisting of: cuprammonium regenerated cellulose, hydrophilic polyvinylidene fluoride (PVDF), composite PVDF, surface modified PVDF, and polyether sulfone. 
     
     
         19 . A method for removing viruses from a liquid Factor VII composition, said composition comprising one or more Factor VII polypeptides, said method comprising subjecting said solution to nanofiltration using a nanofilter having a pore size of 80 nm or less, said nanofilter having a membrane manufactured from one or more materials selected from the group consisting of: cuprammonium regenerated cellulose, hydrophilic polyvinylidene fluoride (PVDF), composite PVDF, surface modified PVDF, and polyether sulfone. 
     
     
         20 . A method for inactivating viruses in a liquid Factor VII composition, said composition comprising one or more Factor VII polypeptides, the method comprising the step of combining said composition with a detergent. 
     
     
         21 . The method according to  claim 20 , wherein the detergent is a octylphenoxy polyethoxyethanol of the formula p-((CH 3 ) 3 CH 2 C(CH 2 ) 2 )—C 6 H 4 —O—(CH 2 CH 2 O) n —H wherein n is in the range of 5-15. 
     
     
         22 . The method according to  claim 20 , wherein the detergent is selected from the list consisting of Triton X-100, Tween®, polysorbate 20, polysorbate 60, and polysorbate 80. 
     
     
         23 . The method according to  claim 20 , wherein the detergent is combined with the liquid Factor VII composition to obtain a concentration of the detergent in the composition in the range of 0.01-0.3% by weight. 
     
     
         24 . The method according to  claim 20 , wherein the detergent is combined with the composition at a temperature of in the range of about 2-12° C. 
     
     
         25 . A method for high-level elimination of active viruses from a liquid Factor VII composition, the method comprising the steps of (i) combining the composition with a detergent, and (ii) subjecting the solution to nanofiltration using a nanofilter having a pore size of 80 nm or less, in any order. 
     
     
         26 . The method according to  claim 25 , wherein the step (i) precedes step (ii).