Production of human IFN- γ (immune) interferon

Claim 2

A process for the production of human immune interferon comprising: (a) providing a viable cell suspension of RPMI 1640 culture medium containing approximately 1.0×10 6 to about 9.0×10 6 human peripheral blood leukocytes per ml; (b) adding a modulator selected from the group consisting of mezerein and 12, 13 phorbol dibutyrate to said cell suspension in an amount of at least 0.7 ηg/ml. of said cell suspension; (c) mixing said modulator in said cell suspension to form a mixture; (d) incubating said mixture under physiological conditions for a period of time sufficient to thoroughly stimulate the cells; (e) mixing Staphylococcal aureus enterotoxin B into said modulated mixture such that a final amount ranging from about 0.01 to about 10.0 μg/ml. of modulater cell suspension is achieved; (f) incubating said modulated mixture under physiological conditions for a period of time sufficient to produce extractable yields of human immune interferon; and (g) separating after incubation the cellular material of the cell suspension from the soluble fraction of the mixture, said soluble fraction containing a substantial portion of the produced human immune interferon and soluble contaminants.

Claim 3

The process in accordance with claim 1, wherein the cell suspension contains approximately 4.0×10 6 to about 7.0×10 6 cells per ml.

Claim 4

The process in accordance with claim 2, wherein the modulator is mezerein.

Claim 5

The process in accordance with claim 2, wherein the modulator is 12, 13 phorbol dibutyrate.

Claim 6

The process according to claims 3 or 4, wherein the modulated mixture is incubated for up to about 3 hours.

Claim 7

The process in accordance with claim 3 or 4, wherein said modulator is added in an amount of between about 1 and about 10 ηg/ml. cell suspension.

Claim 8

The process in accordance with claim 3, wherein said inducer results in a final concentration within said mixture ranging from about 0.1 to about 2.0 μg of inducer/ml. of modulated cell suspension.

Claim 9

The process in accordance with claim 3 or 4, wherein the induced mixture is incubated from about 2 days to about 7 days.

Claim 10

The process in accordance with claim 8, wherein the induced mixture is incubated from about 3 days to about 5 days.

Claim 11

The process in accordance with claim 1, wherein fetal calf serum is added to the mixture in an amount which results in a concentration of fetal calf serum in said mixture of from about 3% to about 15%.

Claim 12

The process according to claim 10, wherein fetal calf serum is added to the induced mixture in an amount which results in a concentration of fetal calf serum in said mixture of from about 4% to about 10%.

Claim 13

The process in accordance with claim 1, wherein the human immune interferon is purified from the contaminants.

Claim 14

A process for the production of human interferon comprising: (a) providing a viable cell suspension of RPMI 1640 culture medium containing approximately 1.0×10 6 to about 9.0×10 6 human peripheral lymphocytes per ml; (b) adding mezerein as a modulator to the cell suspension in an amount of at 0.7 ηg/ml. of cell suspension; (c) mixing said modulator in said cell suspension to form a mixture; (d) incubating said mixture under physiological condition for up to about 3 hours; (e) providing the inducer Staphylococcal aureus enterotoxin B; (f) mixing said inducer in said modulated cell suspension within a container to form an induced mixture wherein said inducer results in a final concentration within said mixture ranging from about 0.01 to about 10.0 μg of inducer/ml of modulated cell suspension; (g) adding fetal calf serum to the mixture in an amount which results in a concentration of fetal calf serum in said mixture of from about 3% to about 15%; (h) incubating said induced mixture under physiological conditions from about 3 to about 7 days; (i) separating after incubation the cellular material of the cell suspension from the soluble fraction of the mixture; said soluble fraction containing a substantial portion of the produced human immune interferon and soluble contaminants; and (j) purifying the human immune interferon from the contaminants.

Claim 15

A process for the production of human interferon comprising: (a) providing a viable cell suspension of RPMI 1640 culture medium containing approximately 1.0×10 6 to about 9.0×10 6 human peripheral lymphocytes per ml; (b) adding mezerein as a modulator to the cell suspension in an amount of at 0.7 ηg/ml. of cell suspension; (c) mixing said modulator in said cell suspension to form a mixture; (d) incubating said mixture under physiological condition for up to about 3 hours; (e) providing the inducer Staphylococcal aureus enterotoxin B; (f) mixing said inducer in said modulated cell suspension within a container to form an induced mixture wherein said inducer results in a final concentration within said mixture ranging from about 0.01 to about 10.0 μg of inducer/ml of modulated cell suspension; (g) adding fetal calf serum to the mixture in an amount which results in a concentration of fetal calf serum in said mixture of from about 3% to about 15%; (h) incubating said induced mixture under physiological conditions from about 3 to about 7 days; (i) separating after incubation the cellular material of the cell suspension from the soluble fraction of the mixture; said soluble fraction containing a substantial portion of the produced human immune interferon and soluble contaminants; and (j) purifying the human immune interferon from the contaminants.