US5362866A
Expired / Lapsed
US
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Oligonucleotide polymeric support system with an oxidation cleavable link
Assignee: Molecular Biosystems Inc
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Filing: 1992-03-17
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Granted: 1994-11-08
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Expires: 2012-03-17
Abstract
Abstract A versatile polymeric support system for the synthesis of oligonucleotides is provided featuring a universal primer which allows chain elongation, in either the 3' or 5' direction, with any currently available DNA or RNA synthesis method, by a process which utilizes oxidatively cleaved primers to facilitate chain elongation and release. The support system is capable of withstanding mildly basic and acidic reaction conditions, while still permitting a convenient and quantitative release, either before or after removal of protecting groups from reactive groups, of synthesized oligonucleotides from a single polymeric support. Removal of the protecting groups before cleavage of the oligomer from the support permits the use of the immobilized oligomer as an affinity hybridization support for both isolating and detecting complementary polynucleic acids.
Inventors
- 👤 Lyle J. Arnold, Jr.
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 16:11
CPC Classifications
C; C07; C07H; C07H21/00; Y; Y02; Y02P; Y02P20/00; Y02P20/50; Y02P20/55
Claims Analysis
Total Claims: 12
Claim 1
Independent Claim
105 words
A process comprising the steps of: (a) providing a compound consisting essentially of a polymeric support and a primer, said compound having the structure: ##STR1## wherein S is a solid support, is a spacer of 1 to 50 atoms which is not cleaved under acidic or basic deprotecting conditions of oligonucleotide synthesis and is attached to said polymeric support, R 8 is a protecting group on an oxidizable substituent and R 7 is a protecting group for --OH; (b) synthesizing an oligonucleotide having base-labile protective groups on the nucleotides onto the O of OR 7 ; and (c) removing the protecting groups with a base.
Claim 2
Dependent Claim
105 words
A process comprising the steps of: (a) providing a compound consisting essentially of a polymeric support and a primer, said compound having the structure: ##STR1## wherein S is a solid support, is a spacer of 1 to 50 atoms which is not cleaved under acidic or basic deprotecting conditions of oligonucleotide synthesis and is attached to said polymeric support, R 8 is a protecting group on an oxidizable substituent and R 7 is a protecting group for --OH; (b) synthesizing an oligonucleotide having base-labile protective groups on the nucleotides onto the O of OR 7 ; and (c) removing the protecting groups with a base.
Claim 3
Dependent Claim
121 words
A process comprising the steps of: (a) providing a compound consisting essentially of a polymeric support and a primer, said compound having the structure: ##STR2## wherein S is a solid support, is a spacer of 1 to 50 atoms which is not cleaved under acidic or basic deprotecting conditions of oligonucleotide synthesis and is attached to said polymeric support, R 8 is a protecting group on an oxidizable substituent, R 7 is a protecting group for --OH, and R 9 are independently selected from the group consisting of H, alkyl, alkoxy and aryl; and (b) synthesizing an oligonucleotide having base-labile protective groups on the nucleotides onto the O of OR 7 ; and (c) removing the protecting groups with a base.
Claim 4
Dependent Claim
121 words
A process comprising the steps of: (a) providing a compound consisting essentially of a polymeric support and a primer, said compound having the structure: ##STR2## wherein S is a solid support, is a spacer of 1 to 50 atoms which is not cleaved under acidic or basic deprotecting conditions of oligonucleotide synthesis and is attached to said polymeric support, R 8 is a protecting group on an oxidizable substituent, R 7 is a protecting group for --OH, and R 9 are independently selected from the group consisting of H, alkyl, alkoxy and aryl; and (b) synthesizing an oligonucleotide having base-labile protective groups on the nucleotides onto the O of OR 7 ; and (c) removing the protecting groups with a base.
Claim 5
Dependent Claim
121 words
A process comprising the steps of: (a) providing a compound consisting essentially of a polymeric support and a primer, said compound having the structure: ##STR3## wherein S is a solid support, is a spacer of 1 to 50 atoms which is not cleaved under acidic or basic deprotecting conditions of oligonucleotide synthesis and is attached to said polymeric support, R 8 is a protecting group on an oxidizable substituent, R 7 is a protecting group for --OH, and is a portion of the cyclic primer that does not directly participate in the cleavage reaction; (b) synthesizing an oligonucleotide having base-labile protective groups on the nucleotides onto the O of OR 7 ; and (c) removing the protecting groups with a base.
Claim 6
Dependent Claim
121 words
A process comprising the steps of: (a) providing a compound consisting essentially of a polymeric support and a primer, said compound having the structure: ##STR3## wherein S is a solid support, is a spacer of 1 to 50 atoms which is not cleaved under acidic or basic deprotecting conditions of oligonucleotide synthesis and is attached to said polymeric support, R 8 is a protecting group on an oxidizable substituent, R 7 is a protecting group for --OH, and is a portion of the cyclic primer that does not directly participate in the cleavage reaction; (b) synthesizing an oligonucleotide having base-labile protective groups on the nucleotides onto the O of OR 7 ; and (c) removing the protecting groups with a base.
Claim 7
Dependent Claim
136 words
A process comprising the steps of: (a) providing a compound consisting essentially of a polymeric support and a primer, said compound having the structure: ##STR4## wherein S is a solid support, is a spacer of 1 to 50 atoms which is not cleaved under acidic or basic deprotecting conditions of oligonucleotide synthesis and is attached to said polymeric support, R 8 is a protecting group on an oxidizable substituent, R 7 is a protecting group for --OH, R 9 are independently selected from the group consisting of H, alkyl, alkoxy and aryl and is the portion of the cyclic primer that does not directly participate in the cleavage reaction; (b) synthesizing an oligonucleotide having base-labile protective groups on the nucleotides onto the O of OR 7 ; and (c) removing the protecting groups with a base.
Claim 8
Dependent Claim
136 words
A process comprising the steps of: (a) providing a compound consisting essentially of a polymeric support and a primer, said compound having the structure: ##STR4## wherein S is a solid support, is a spacer of 1 to 50 atoms which is not cleaved under acidic or basic deprotecting conditions of oligonucleotide synthesis and is attached to said polymeric support, R 8 is a protecting group on an oxidizable substituent, R 7 is a protecting group for --OH, R 9 are independently selected from the group consisting of H, alkyl, alkoxy and aryl and is the portion of the cyclic primer that does not directly participate in the cleavage reaction; (b) synthesizing an oligonucleotide having base-labile protective groups on the nucleotides onto the O of OR 7 ; and (c) removing the protecting groups with a base.
Claim 9
Dependent Claim
46 words
The process of any one of claims 1-4 further comprising the steps of: (d) oxidizing at least the oxidizable substituent OR 8 that is gamma to the oligonucleotide attachment site; (e) cleaving the oligonucleotide from the primer with a second base; and (f) recovering the oligonucleotide.
Claim 10
Dependent Claim
25 words
The process of any one of claims 1-4 further comprising the step of: (d) hybridizing the oligonucleotide to a complementary oligonucleotide to form a duplex.
Claim 11
Dependent Claim
19 words
The process of claim 6 further comprising the step of: (e) detecting the complementary oligonucleotide hybridized to the oligonucleotide.
Claim 12
Dependent Claim
43 words
The process of claim 6 further comprising the steps of: (e) oxidizing at least the oxidizable substituent OR 8 that is gamma to the oligonucleotide attachment site; (f) cleaving the oligonucleotide from the primer with a second base; and (g) recovering the duplex.
References Cited (Backward Citations)
US4500707A
Cited
US4959463A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1997-11-07 | 1998-05-09 | 1998-11-08 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2001-11-07 | 2002-05-09 | 2002-11-08 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2005-11-07 | 2006-05-09 | 2006-11-08 | $7700.00 | Window Closed / Lapsed |