VIRAL INACTIVATION USING IMPROVED SOLVENT-DETERGENT METHOD
The present specification discloses methods of inactivating a lipid-coat containing virus and proteins essentially free of a lipid-coat containing virus obtained from such methods.
1 - 85 . (canceled)
86 . A method of inactivating a lipid-coat containing virus, the method comprising:
(a) admixing tri(n-butyl) phosphate, polyoxyethylene octyl phenyl, and polysorbate 80 sorbitan with a cell culture supernatant comprising recombinant Factor VIII having a first activity, thereby forming a mixture, wherein the admixing is conducted at a temperature from 2° C. to 6° C., and wherein the final concentration of tri(n-butyl) phosphate in the mixture is from about 0.1% (v/v) to about 5.0% (v/v), the final concentration of polyoxyethylene octyl phenyl in the mixture is from about 0.5% (v/v) to about 10.0% (v/v), and the final concentration of polysorbate 80 sorbitan in the mixture is from about 0.1% (v/v) to about 5.0% (v/v); and
(b) incubating the mixture for no more than about 120 minutes at a temperature of from 2° C. to 6° C., thereby forming an incubated mixture, essentially free of viable lipid-coat containing virus, and
wherein Factor VIII in the incubated mixture has a second activity of at least 25% of the first activity.
87 . The method of claim 86 , wherein the final concentration of tri(n-butyl) phosphate is from about 0.1% (v/v) to about 0.7%, the final concentration of polyoxyethylene octyl phenyl is from about 0.6% (v/v) to about 1.4% (v/v), and the final concentration of polysorbate 80 sorbitan is from about 0.1% (v/v) to about 0.7%
88 . The method of claim 86 , wherein the final concentration of tri(n-butyl) phosphate is from about 0.1% (v/v) to about 0.5%, the final concentration of polyoxyethylene octyl phenyl is from about 0.7% (v/v) to about 1.3% (v/v), and the final concentration of polysorbate 80 sorbitan is from about 0.1% (v/v) to about 0.5%
89 . The method of claim 86 , wherein the final concentration of tri(n-butyl) phosphate is from about 0.2% (v/v) to about 0.4%, the final concentration of polyoxyethylene octyl phenyl is from about 0.8% (v/v) to about 1.2% (v/v), and the final concentration of polysorbate 80 sorbitan is from about 0.2% (v/v) to about 0.4%.
90 . The method of claim 86 , wherein the final concentration of tri(n-butyl) phosphate is about 0.3% (v/v), the final concentration of polyoxyethylene octyl phenyl is about 1.0% (v/v), and the final concentration of polysorbate 80 sorbitan is about 0.3% (v/v).
91 . The method of claim 86 , wherein less than 1% of the Factor VIII in the incubated mixture is aggregated.
92 . The method of claim 86 , wherein Factor VIII in the incubated mixture has a second activity of at least 50% of the first activity.
93 . The method of claim 86 , wherein the mixture is incubated in step (b) for no more than about 60 minutes.
94 . The method of claim 86 , wherein the mixture is incubated in step (b) for no more than about 30 minutes.
95 . The method of claim 86 , wherein the mixture is incubated in step (b) for no more than about 10 minutes.
96 . The method of claim 86 , wherein both steps (a) and (b) are performed at a temperature of from 2° C. to 4° C.
97 . The method of claim 86 , wherein both steps (a) and (b) are performed at a temperature of about 4° C.
98 . The method of claim 86 , wherein the method is capable of reducing the viral infectivity of a herpesviridae virus in the cell culture supernatant by at least 4 log.
99 . The method of claim 86 , wherein the method is capable of reducing the viral infectivity of a flaviviridae virus in the cell culture supernatant by at least 4 log.
100 . The method of claim 86 , wherein the method is capable of reducing the viral infectivity of pseudorabies virus (PRV) in the fluid by at least 4 log.
101 . A composition comprising Factor VIII that is essentially free of a lipid-coat containing virus prepared according to the method of claim 86 .
102 . A method of inactivating a lipid-coat containing virus, the method comprising:
(a) admixing an alkylphosphate and at least two non-ionic surfactants with a fluid comprising a blood protein having a first activity, thereby forming a mixture; and
(b) incubating the mixture for no more than about 120 minutes, thereby forming an incubated mixture,
wherein the incubated mixture is essentially free of viable lipid-coat containing virus, and
wherein the blood protein in the incubated mixture has a second activity of at least 25% of the first activity.
103 . A composition comprising a blood factor that is essentially free of a lipid-coat containing virus prepared according to the method of claim 102 .