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US 4,376,821 AGranted
Production of human IFN-gamma (immune) interferon
AssigneeMeloy Laboratories, Inc.
InventorIrwin A. Braude
Life of this patent
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Abstract
A process is disclosed for producing human immune interferon with cell cultures of human peripheral blood leukocytes modulated with Mezerein or 12, 13 phorbol dibutyrate and induced with the calcium ionophore A-23187. After cultures are incubated for 12 hours to about 7 days, crude interferon on the order of 10.sup.3 units of interferon/ml. is produced.
Claims15
A process for the production of human immune interferon comprising:
- (a) providing a viable cell suspension containing approximately 1.0.times.10.sup.6 to about 9.0.times.10.sup.6 human peripheral blood leukocytes per ml;
- (b) adding a modulator to said cell suspension said modulator being selected from the group consisting of mezerein and 12,13 phorbol dibutyrate in an amount of at least 0.7 .eta.g/ml. of said cell suspension;
- (c) adding calcium ionophore A-23187 to said cell suspension such that a final amount ranging from about 0.1 to about 10.0 .mu.g/ml of cell suspension is achieved;
- (d) mixing said modulator and said inducer in said cell suspension to form a mixture;
- (e) adding a protein supplement in an amount which results in a concentration of supplement sufficient to keep the cells alive;
- (f) incubating said 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.
5 more elements
A process for the production of human interferon comprising:
- (a) providing a viable cell suspension containing approximately 1.0.times.10.sup.6 to about 9.0.times.10.sup.6 human peripheral leukocytes per ml;
- (b) adding mezerein as a modulator to the cell suspension in an amount of at 0.7 .eta.g/ml. of cell suspension.
- (c) incubating said mixture under physiological condition for up to about 3 hours;
- (d) providing as the inducer the calcium ionophore A-23187;
- (e) mixing said modular and inducer in said cell suspension within a container to form an induced mixture wherein said inducer results in a final concentration within said mixture ranging from about 0.1 to about 10.0 .mu.g of inducer/ml of cell suspension.
- (f) incubating said induced mixture under physiological conditions from about 12 hours to about 7 days;
- (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;
- and
- (h) purifying the human immune interferon from the contaminants.
7 more elements
Dependencies are inferred from the claim text.