Back to results
US 4,376,822 AGranted
Production of human IFN- .gamma. (immune) interferon
AssigneeMeloy Laboratories, Inc.
InventorIrwin A. Braude
Life of this patent
Estimating the term…
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 Staphylococcal aureus enterotoxin B. After cultures are incubated for 3 to 7 days crude interferon on the order of 10.sup.3 units of interferon/ml. is produced.
Claims13
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.times.10.sup.6 to about 9.0.times.10.sup.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 .eta.g/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 .mu.g/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.
5 more elements
A process for the production of human interferon comprising:
- (a) providing a viable cell suspension of RPMI 1640 culture medium containing approximately 1.0.times.10.sup.6 to about 9.0.times.10.sup.6 human peripheral lymphocytes 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) 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 .mu.g 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.
9 more elements
Dependencies are inferred from the claim text.