US6203975B1
Expired / Lapsed
US
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Adenovirus and method of use thereof
Assignee: University of Pennsylvania
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Filing: 1999-10-21
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Granted: 2001-03-20
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Expires: 2019-10-21
Abstract
Abstract A recombinant adenovirus and a method for producing the virus are provided which utilize a recombinant shuttle vector comprising adenovirus DNA sequence for the 5′ and 3′ cis-elements necessary for replication and virion encapsidation in the absence of sequence encoding viral genes and a selected minigene linked thereto, and a helper adenovirus comprising sufficient adenovirus gene sequences necessary for a productive viral infection. Desirably the helper gene is crippled by modifications to its 5′ packaging sequences, which facilitates purification of the viral particle from the helper virus.
Inventors
- 👤 James M. Wilson
- 👤 Krishna J. Fisher
- 👤 Shu-Jen Chen
- 👤 Matthew Weitzman
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 19:58
CPC Classifications
C; C12; C12N; C12N7/00; C12N15/00; C12N15/09; C12N15/63; C12N15/79; C12N15/85; C12N15/86; A; A61; A61K; A61K48/00; C12N2710/00; C12N2710/00011; C12N2710/10011; C12N2710/10311; C12N2710/10341; C12N2710/10343; C12N2810/00; C12N2810/50; C12N2810/60; C12N2810/6009; C12N2810/6018; C12N2840/00; C12N2840/44
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
156 words
A method for producing an AdΔ virus which comprises the steps of: (a) introducing into a selected host cell: (i) a recombinant shuttle vector which comprises adenovirus nucleic acid sequences and a minigene, wherein said adenovirus sequences are of a first serotype and consist of the adenovirus 5′ and 3′ cis-elements necessary for replication and virion encapsidation; and wherein said minigene comprises a selected gene operatively linked to regulatory sequences which direct expression of said selected gene in a target cell, said adenovirus cis-elements flanking said minigene, and (ii) a helper virus comprising adenovirus gene sequences necessary for a productive viral infection and encoding an adenoviral capsid of a second serotype; (b) culturing said host cell containing the shuttle vector and the helper virus so as to permit the formation of an AdΔ virus comprising the adenovirus sequences and minigene of (i) in an adenoviral capsid of a second serotype which differs from said first serotype.
Claim 2
Independent Claim
156 words
A method for producing an AdΔ virus which comprises the steps of: (a) introducing into a selected host cell: (i) a recombinant shuttle vector which comprises adenovirus nucleic acid sequences and a minigene, wherein said adenovirus sequences are of a first serotype and consist of the adenovirus 5′ and 3′ cis-elements necessary for replication and virion encapsidation; and wherein said minigene comprises a selected gene operatively linked to regulatory sequences which direct expression of said selected gene in a target cell, said adenovirus cis-elements flanking said minigene, and (ii) a helper virus comprising adenovirus gene sequences necessary for a productive viral infection and encoding an adenoviral capsid of a second serotype; (b) culturing said host cell containing the shuttle vector and the helper virus so as to permit the formation of an AdΔ virus comprising the adenovirus sequences and minigene of (i) in an adenoviral capsid of a second serotype which differs from said first serotype.
Claim 3
Dependent Claim
32 words
The method according to claim 1 , wherein the first and second serotypes are individually selected from an adenovirus serotype of the group consisting of 2, 4, 5, 7, 12 and 40.
Claim 4
Dependent Claim
25 words
The method according to claim 1 , further comprising the steps of isolating and purifying the AdΔ virus from said host cell or cell culture.
Claim 5
Independent Claim
173 words
A method for producing an AdΔ virus which comprises the steps of: (a) introducing into a selected host cell (i) a recombinant shuttle vector which comprises adenovirus nucleic acid sequences and a minigene, wherein said adenovirus sequences are of a first serotype and consist of the adenovirus 5′ and 3′ cis-elements necessary for replication and virion encapsidation; and wherein said minigene comprises a selected gene operatively linked to regulatory sequences which direct expression of said selected gene in a target cell, said adenovirus cis-elements flanking said minigene; and (ii) a helper virus comprising adenovirus gene sequences of a serotype which differ s from the serotype of the adenovirus sequences of (i); (b) culturing said host cell containing the shuttle vector and the helper virus, wherein the helper virus and the host cell provide the adenovirus genes necessary for a productive viral infection and which permit the formation of an AdΔ virus comprising the adenovirus sequences and minigene of (i) in an adenoviral capsid of a second serotype which differs from said first serotype.
Claim 6
Independent Claim
173 words
A method for producing an AdΔ virus which comprises the steps of: (a) introducing into a selected host cell (i) a recombinant shuttle vector which comprises adenovirus nucleic acid sequences and a minigene, wherein said adenovirus sequences are of a first serotype and consist of the adenovirus 5′ and 3′ cis-elements necessary for replication and virion encapsidation; and wherein said minigene comprises a selected gene operatively linked to regulatory sequences which direct expression of said selected gene in a target cell, said adenovirus cis-elements flanking said minigene; and (ii) a helper virus comprising adenovirus gene sequences of a serotype which differ s from the serotype of the adenovirus sequences of (i); (b) culturing said host cell containing the shuttle vector and the helper virus, wherein the helper virus and the host cell provide the adenovirus genes necessary for a productive viral infection and which permit the formation of an AdΔ virus comprising the adenovirus sequences and minigene of (i) in an adenoviral capsid of a second serotype which differs from said first serotype.
Claim 7
Dependent Claim
32 words
The method according to claim 4 , wherein the first and second serotypes are individually selected from an adenovirus serotype of the group consisting of 2, 4, 5, 7, 12 and 40.
Claim 8
Dependent Claim
25 words
The method according to claim 4 , further comprising the steps of isolating and purifying the AdΔ virus from said host cell or cell culture.
Claim 9
Dependent Claim
92 words
A recombinant AdΔ virus comprising: (a) adenovirus nucleic acid sequences and a minigene, wherein said adenovirus sequences are of a first serotype and consist of the adenovirus 5′ and 3′ cis-elements necessary for replication and virion encapsidation; and wherein said minigene comprises a selected gene operatively linked to regulatory sequences which direct expression of said selected gene in a target cell, said adenovirus cis-elements flanking said minigene; and (b) a capsid of a second adenovirus serotype which differs from said first serotype and which encapsidates said adenovirus sequences and minigene of (a).
Claim 10
Dependent Claim
92 words
A recombinant AdΔ virus comprising: (a) adenovirus nucleic acid sequences and a minigene, wherein said adenovirus sequences are of a first serotype and consist of the adenovirus 5′ and 3′ cis-elements necessary for replication and virion encapsidation; and wherein said minigene comprises a selected gene operatively linked to regulatory sequences which direct expression of said selected gene in a target cell, said adenovirus cis-elements flanking said minigene; and (b) a capsid of a second adenovirus serotype which differs from said first serotype and which encapsidates said adenovirus sequences and minigene of (a).
Claim 11
Dependent Claim
32 words
The virus according to claim 7 , wherein the first and second serotypes are individually selected from an adenovirus serotype of the group consisting of 2, 4, 5, 7, 12 and 40.
Claim 12
Independent Claim
105 words
A method for producing a pseudotyped AdΔ virus comprising the steps of: culturing, under conditions which permit the formation of a pseudotyped AdΔ virus, a host cell containing: (i) adenovirus 5′ and 3′ cis-elements necessary for replication and virion encapsidation and a transgene under the control of regulatory elements which control expression thereof in a host cell, wherein said adenovirus 5′ and 3′ cis-elements flank said transgene and are of a first adenovirus serotype; (ii) an adenovirus capsid, wherein said adenovirus capsid is of a second adenovirus serotype, which differs from said first adenovirus serotype; and (iii) adenovirus sequences necessary for a productive viral infection.
Claim 13
Independent Claim
105 words
A method for producing a pseudotyped AdΔ virus comprising the steps of: culturing, under conditions which permit the formation of a pseudotyped AdΔ virus, a host cell containing: (i) adenovirus 5′ and 3′ cis-elements necessary for replication and virion encapsidation and a transgene under the control of regulatory elements which control expression thereof in a host cell, wherein said adenovirus 5′ and 3′ cis-elements flank said transgene and are of a first adenovirus serotype; (ii) an adenovirus capsid, wherein said adenovirus capsid is of a second adenovirus serotype, which differs from said first adenovirus serotype; and (iii) adenovirus sequences necessary for a productive viral infection.
Claim 14
Dependent Claim
32 words
The method according to claim 9 , wherein the first and second serotypes are individually selected from an adenovirus serotype of the group consisting of 2, 4, 5, 7, 12, and 40.
Claim 15
Dependent Claim
23 words
The method according to claim 9 , further comprising the steps of isolating the AdΔ virus from said host cell or cell culture.
Claim 16
Dependent Claim
23 words
The method according to claim 9 , further comprising the steps of purifying the AdΔ virus from said host cell or cell culture.
References Cited (Backward Citations)
US5166320A
Cited
US5240846A
Cited
WO1991018088A1
Cited
WO1993003769A1
Cited
US5252479A
Cited
WO1993019191A1
Cited
WO1994008026A1
Cited
WO1994012649A2
Cited
US5882877A
Cited
WO1994017832A1
Cited
WO1994028152A1
Cited
WO1994028938A1
Cited
WO1995000655A1
Cited
WO1995002697A1
Cited
US5543328A
Cited
WO1995027071A2
Cited
WO1995029993A1
Cited
WO1995034671A1
Cited
WO1996013596A1
Cited
US5856152A
Cited
US5871982A
Cited
WO1996014061A1
Cited
WO1996018418A1
Cited
US5652224A
Cited
US5872154A
Cited
US5756283A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2004-03-19 | 2004-09-18 | 2005-03-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2008-03-19 | 2008-09-18 | 2009-03-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2012-03-19 | 2012-09-18 | 2013-03-20 | $7700.00 | Window Closed / Lapsed |