US4564593A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Messenger RNA, production and use thereof
Assignee: Takeda Pharmaceutical Co Ltd
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Filing: 1982-12-22
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Granted: 1986-01-14
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Expires: 2002-12-22
Abstract
Abstract The present invention provides human Interleukin 2-encording messenger RNA which is free of human cells, a method of producing the same from the cells which are capable of producing human Interleukin 2 and a method of producing human Interleukin 2 using the same in an in vitro protein synthesis system.
Inventors
- 👤 Kyozo Tsukamoto
- 👤 Shuji Hinuma
- 👤 Haruo Onda
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 06:52
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/54; C07K14/55
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
28 words
A culture of cells containing exogenous messenger RNA isolated from human cells which has a sedimentation coefficient of 85 to 155 and which codes for human Interleukin 2.
Claim 2
Dependent Claim
28 words
A culture of cells containing exogenous messenger RNA isolated from human cells which has a sedimentation coefficient of 85 to 155 and which codes for human Interleukin 2.
Claim 3
Dependent Claim
24 words
Substantially pure messenger RNA isolated from human cells, which has a sedimentation coefficient of 8S to 15S and which codes for human Interleukin 2.
Claim 4
Dependent Claim
24 words
Substantially pure messenger RNA isolated from human cells, which has a sedimentation coefficient of 8S to 15S and which codes for human Interleukin 2.
Claim 5
Dependent Claim
22 words
Substantially pure messenger RNA according to claim 1, wherein the human Interleukin 2 is human T-cell growth factor or human co-stimulating factor.
Claim 6
Independent Claim
136 words
A method of producing messenger RNA isolated from human cells, the messenger RNA having a sedimentation coefficient of 8S to 15S and coding for human Interleukin 2, which comprises growing human Interleukin 2-producing lymphocytic cells isolated from human peripheral blood, in a medium for animal cell culture containing lectin and phorbol ester as an inducer, at 30° to 40° C., by spinner culture, for about 5 to 80 hours, to cause formation and accumulation of the messenger RNA in the cells, harvesting the grown cells by centrifugation, homogenzing the resulting cells in the presence of buffer containing thiocyanate and mercapto-alkanol, centrifuging the homogenate in the presence of cesium chloride, precipitating RNA from the resulting supernatant by adding sodium chloride and lower alkanol, and purifying the RNA by oligo (dT)-cellulose column chromatography and sucrose density gradient centrifugation.
Claim 7
Independent Claim
136 words
A method of producing messenger RNA isolated from human cells, the messenger RNA having a sedimentation coefficient of 8S to 15S and coding for human Interleukin 2, which comprises growing human Interleukin 2-producing lymphocytic cells isolated from human peripheral blood, in a medium for animal cell culture containing lectin and phorbol ester as an inducer, at 30° to 40° C., by spinner culture, for about 5 to 80 hours, to cause formation and accumulation of the messenger RNA in the cells, harvesting the grown cells by centrifugation, homogenzing the resulting cells in the presence of buffer containing thiocyanate and mercapto-alkanol, centrifuging the homogenate in the presence of cesium chloride, precipitating RNA from the resulting supernatant by adding sodium chloride and lower alkanol, and purifying the RNA by oligo (dT)-cellulose column chromatography and sucrose density gradient centrifugation.
Claim 8
Dependent Claim
35 words
A method according to claim 3, wherein the inducer comprises concanavalin A as lectin and 12-O-tetradecanoylphorbol 13-acetate as phorbol ester in the concentrations between about 5 to 80 μg/ml and 1 to 50 ng/ml, respectively.
Claim 9
Dependent Claim
38 words
The process of producing cDNA coding for human Interleukin 2 comprising contacting messenger RNA isolated from human cells and coding for human Interleukin 2 with reverse transcriptase for a time and under conditions sufficient to form said cDNA.
Claim 10
Dependent Claim
38 words
The process of producing cDNA coding for human Interleukin 2 comprising contacting messenger RNA isolated from human cells and coding for human Interleukin 2 with reverse transcriptase for a time and under conditions sufficient to form said cDNA.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1989-01-13 | 1989-07-15 | 1990-01-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-01-13 | 1993-07-15 | 1994-01-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-01-13 | 1997-07-15 | 1998-01-14 | $7700.00 | Window Closed / Lapsed |