US5024939A
Expired / Lapsed
US
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Transient expression system for producing recombinant protein
Assignee: Genentech Inc
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Filing: 1987-09-25
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Granted: 1991-06-18
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Expires: 2007-09-25
Abstract
Abstract A method is described for transient production of a desired heterologous protein comprising: transfecting a eukaryotic host cell with a vector producing a trans-activating protein; transfecting the eukaryotic host cell with an expression vector comprising a stabilizing sequence downstream of a promoter and upstream of a DNA encoding the desired heterologous protein and a polyadenylation sequence downstream of which is a transcription terminatein site; culturing the transfected eukaryotic host cell under conditions favorable for production of said desired heterologous protein; and, recovering the desired protein in useful amounts within about one day to about fourteen days of transfection.
Inventors
- 👤 Cornelia M. Gorman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 15:58
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/14; C12N9/48; C12N9/50; C12N9/64; C12N9/6421; C12N9/6424; C12N9/6456; C12N9/6459; C07; C07K; C07K14/00; C07K14/435; C07K14/575; C07K14/61; C07K14/64; C07K14/745; C07K14/755; C12N15/00; C12N15/09; C12N15/63; C12N15/67; C12N15/79; C12N15/85; C12Y; C12Y304/00; C12Y304/21; C12Y304/21069
Claims Analysis
Total Claims: 30
Claim 1
Independent Claim
76 words
A method for the enhanced transient expression of a desired protein in a host kidney cell comprising: a) providing a host kidney cell which expresses an adenovirus trans-activating protein; b) transfecting said host kidney cell with a vector comprising DNA encoding the desired protein under the control of a cytomegalovirus promoter; and, c) culturing said host kidney cell for a period comprising 1 to 14 days under conditions favorable for transient production of the desired protein.
Claim 2
Independent Claim
76 words
A method for the enhanced transient expression of a desired protein in a host kidney cell comprising: a) providing a host kidney cell which expresses an adenovirus trans-activating protein; b) transfecting said host kidney cell with a vector comprising DNA encoding the desired protein under the control of a cytomegalovirus promoter; and, c) culturing said host kidney cell for a period comprising 1 to 14 days under conditions favorable for transient production of the desired protein.
Claim 3
Dependent Claim
12 words
The method of claim 1 wherein said adenovirus trans-activating protein is Elb.
Claim 4
Dependent Claim
12 words
The method of claim 1 further comprising the presence of T antigen.
Claim 5
Dependent Claim
12 words
The method of claim 1 wherein said adenovirus trans-activating protein is Ela.
Claim 6
Dependent Claim
17 words
The method of claim 1 wherein said adenovirus trans-activating protein is a mixture of Ela and Elb.
Claim 7
Dependent Claim
18 words
The method of claim 1 wherein said cytomegalovirus promoter is from the immediate early gene of human cytomegalovirus.
Claim 8
Dependent Claim
16 words
The method of claim 1 wherein said host kidney cell is a human embryonic kidney cell.
Claim 9
Dependent Claim
13 words
The method of claim 7 wherein said human embryonic kidney cell is 293.
Claim 10
Dependent Claim
14 words
The method of claim 7 wherein said human embryonic kidney host cell is JW2.
Claim 11
Dependent Claim
27 words
The method of claim 7 which includes under step b) the additional step b)1) transfecting said host kidney cell with a vector producing a translational control effector.
Claim 12
Dependent Claim
13 words
The method of claim 10 wherein the translational control effector is VA RNA.
Claim 13
Dependent Claim
20 words
The method of claim 1 wherein said step b) further comprises downstream from said cytomegalovirus promoter a stabilizing DNA sequence.
Claim 14
Dependent Claim
39 words
A host kidney cell wherein said host kidney cell endogenously produces an adenovirus trans-activating protein and is transformed by transfecting said host kidney cell with a vector comprising DNA encoding said protein under the control of a cytomegalovirus promoter.
Claim 15
Dependent Claim
39 words
A host kidney cell wherein said host kidney cell endogenously produces an adenovirus trans-activating protein and is transformed by transfecting said host kidney cell with a vector comprising DNA encoding said protein under the control of a cytomegalovirus promoter.
Claim 16
Dependent Claim
14 words
The host kidney cell of claim 13 that is a human embryonic kidney cell.
Claim 17
Dependent Claim
14 words
The host human embryonic kidney cell of claim 14 that is derived from JW2.
Claim 18
Dependent Claim
14 words
The host human embryonic kidney cell of claim 14 that is derived from 293.
Claim 19
Dependent Claim
53 words
In a method for production of a host kidney cell capable of production of a desired heterologous protein, wherein said host kidney cell endogenously produces an adenovirus trans-activating protein, the improvement comprising transfecting said host kidney cell with a vector comprising DNA encoding said desired protein under the control of a cytomegalovirus promoter.
Claim 20
Dependent Claim
53 words
In a method for production of a host kidney cell capable of production of a desired heterologous protein, wherein said host kidney cell endogenously produces an adenovirus trans-activating protein, the improvement comprising transfecting said host kidney cell with a vector comprising DNA encoding said desired protein under the control of a cytomegalovirus promoter.
Claim 21
Dependent Claim
12 words
The method of claim 17 wherein said adenovirus trans-activating protein is Ela.
Claim 22
Dependent Claim
12 words
The method of claim 17 wherein said adenovirus trans-acting protein is Elb.
Claim 23
Dependent Claim
17 words
The method of claim 17 wherein said adenovirus trans-activating protein is a mixture of Ela and Elb.
Claim 24
Dependent Claim
15 words
The method of claim 17 wherein said kidney cell is a human embryonic kidney cell.
Claim 25
Dependent Claim
16 words
The method of claim 21 wherein said human embryonic kidney cell is the 293 cell line.
Claim 26
Dependent Claim
16 words
The method of claim 21 wherein said human embryonic kidney cell is the JW2 cell line.
Claim 27
Dependent Claim
14 words
The method of claim 17 wherein said cytomegalovirus promoter is an immediate early promoter.
Claim 28
Dependent Claim
19 words
The method of claim 24 wherein said immediate early promoter is the human immediate early promoter of human cytomegalovirus.
Claim 29
Dependent Claim
20 words
The method of claim 17 further comprising the additional step of transfecting with a vector producing a translational control effector.
Claim 30
Dependent Claim
13 words
The method of claim 26 wherein said translational control effector is VA RNA.
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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) | 1994-06-17 | 1994-12-17 | 1995-06-18 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1998-06-17 | 1998-12-17 | 1999-06-18 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2002-06-17 | 2002-12-17 | 2003-06-18 | $7700.00 | Window Closed / Lapsed |