Patent US4777129A successfully refreshed from Google Patents.
US4777129A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Nucleic acid probe detectable by specific nucleic acid binding protein
Assignee: MILES INC. A CORP. OF INDIANA
•
Filing: 1984-10-19
•
Granted: 1988-10-11
•
Expires: 2004-10-19
Abstract
Abstract A nucleic acid detection probe comprising a hybridizable single stranded portion of nucleic acid connected with a non-hybridizable, single or double stranded nucleic acid portion, the non-hybridizable portion preferably including a recognition site for binding by a particular protein. Such recognition site can be a region of singly or doubly stranded nucleic acid specific for a particular nucleic acid binding protein such as lac repressor protein or can be a modified nucleic acid region such as a unique antigenic determinant introduced by interaction of the region with a modifier compound such as an intercalating agent or a platinum-containing ligand. The probe-binding protein can be labeled for ease of detection and in the case of an antigenic determinant binding site can be labeled antibody.
Inventors
- 👤 Nanibhushan Dattagupta
- 👤 Peter M. M. Rae
- 👤 William J. Knowles
- 👤 Donald M. Crothers
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-24 04:47
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6876; Y; Y10; Y10S; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/811
Claims Analysis
Total Claims: 49
Claim 1
Independent Claim
85 words
A nucleic acid detection probe comprising a hybridizable single stranded portion of nucleic acid connected with a non-hybridizable, single stranded nucleic acid portion, the non-hybridizable portion including a recognition site for binding by a particular protein, wherein the non-hybridizable portion has been modified to create the protein recognition site and wherein the modification is effected by connecting to said non-hybridizable portion an intercalating agent or wherein the modification is effected by contact with a modifier compound which introduces an antigenic determinant into the non-hybridizable portion.
Claim 2
Dependent Claim
85 words
A nucleic acid detection probe comprising a hybridizable single stranded portion of nucleic acid connected with a non-hybridizable, single stranded nucleic acid portion, the non-hybridizable portion including a recognition site for binding by a particular protein, wherein the non-hybridizable portion has been modified to create the protein recognition site and wherein the modification is effected by connecting to said non-hybridizable portion an intercalating agent or wherein the modification is effected by contact with a modifier compound which introduces an antigenic determinant into the non-hybridizable portion.
Claim 3
Dependent Claim
18 words
A detection probe according to claim 1, wherein the hybridizable portion is covalently linked to the non-hybridizable portion.
Claim 4
Dependent Claim
22 words
A detection probe according to claim 1, wherein the hybridizable portion is complementary to the genomic sequence responsible for sickle cell anemia.
Claim 5
Dependent Claim
22 words
A detection probe according to claim 1, wherein the non-hybridizable portion comprises single stranded nucleic acid photochemically linked with an intercalating agent.
Claim 6
Dependent Claim
27 words
A detection probe according to claim 1, wherein the modifier compound is an intercalating agent selected from the group consisting of acridine dyes, phenazines, phenothiazines, and quinolines.
Claim 7
Dependent Claim
20 words
A detection probe according to claim 1, including the particular protein bound to the protein-specific site of the non-hybridizable portion.
Claim 8
Dependent Claim
13 words
A detection probe according to claim 6, wherein the protein carries a label.
Claim 9
Dependent Claim
30 words
A detection probe according to claim 7, wherein the label is selected from the group consisting of a fluorescer, a chromophore, a luminescer, a specifically bindable ligand and a radioisotope.
Claim 10
Dependent Claim
69 words
A detection probe according to claim 1, including a labeled from of the particular protein bound to the protein-specific site of the non-hybridizable portion, wherein the labeled protein comprises a protein selected from the group consisting of an antibody, and an antibody fragment and a label selected from the group consisting of an enzymatically active group, a fluorescer, a chromophore, a liminescer, a specifically bindable ligand and a radioisotope.
Claim 11
Independent Claim
164 words
A method for detecting a particular polynucleotide sequence in a test medium containing single stranded nucleic acids, comprising the steps of: (a) combining a test medium with a nucleic acid detection probe comprising at least one hybridizable single stranded base sequence to be detected and a non-hybridizable double-stranded portion having a recognition site for binding by a particular protein, under conditions favorable to hybridization between the sequence to be detected and the complementary hybridizable sequence in the probe, wherein the non-hybridizable portion has been modified to create the protein recognition site, wherein the modification is effected by connecting to said non-hybridizable portion an intercalating agent or a platinum-containing ligand or the modification is effected by introducing an antigenic determinant into the non-hybridizable portion, (b) separating hybridized probe from unhybridized probe, and (c) adding to the separated hybridized probe, the particular protein which binds the recognition site on the non-hybridizable portion of the probe and determining the protein which becomes bound to the solid support.
Claim 12
Independent Claim
164 words
A method for detecting a particular polynucleotide sequence in a test medium containing single stranded nucleic acids, comprising the steps of: (a) combining a test medium with a nucleic acid detection probe comprising at least one hybridizable single stranded base sequence to be detected and a non-hybridizable double-stranded portion having a recognition site for binding by a particular protein, under conditions favorable to hybridization between the sequence to be detected and the complementary hybridizable sequence in the probe, wherein the non-hybridizable portion has been modified to create the protein recognition site, wherein the modification is effected by connecting to said non-hybridizable portion an intercalating agent or a platinum-containing ligand or the modification is effected by introducing an antigenic determinant into the non-hybridizable portion, (b) separating hybridized probe from unhybridized probe, and (c) adding to the separated hybridized probe, the particular protein which binds the recognition site on the non-hybridizable portion of the probe and determining the protein which becomes bound to the solid support.
Claim 13
Dependent Claim
40 words
A method according to claim 10 which is a solid phase hybridization technique, wherein the single stranded nucleic acids from the test medium are immobilized on a solid support and wherein the protein associated with the solid support is determined.
Claim 14
Dependent Claim
76 words
A method according to claim 10 which is a solid phase sandwich hybridization technique, wherein the test medium is combined with first and second nucleic acid probes each comprising at least one hybridizable single stranded base sequence which is substantially complementary to a mutually exclusive portion of the sequence to be detected and wherein one of the probes is immobilized on a solid support and the other has said non-hybridizable portion having said protein recognition site.
Claim 15
Dependent Claim
21 words
A method according to claim 10, wherein the non-hybridizable portion comprises single stranded nucleic acid photochemically linked with an intercalating agent.
Claim 16
Dependent Claim
25 words
A method according to claim 10, wherein the non-hybridizable portion comprises double stranded nucleic acid in the form of intercalating complexes with an intercalating agent.
Claim 17
Dependent Claim
26 words
A method according to claim 14, wherein the intercalating agent is covalently linked to at least one of the two strands of the double stranded portion.
Claim 18
Dependent Claim
38 words
A method according to claim 10, wherein the modification is effected by contact with a modifier compound, said modifier compound being an intercalating agent selected from the group consisting of acaridine dyes, phenanthridines, phenazines, furocouramarins, phenothiazines and quinolines.
Claim 19
Dependent Claim
18 words
A method according to claim 10, wherein the non-hybridizable portion has been modified by contact with a salt.
Claim 20
Dependent Claim
13 words
A method according to claim 10, wherein the particular protein carries a label.
Claim 21
Dependent Claim
33 words
A method according to claim 18, wherein the label is selected from the group consisting of an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand and a radioisotope.
Claim 22
Dependent Claim
43 words
A method according to claim 10, wherein the protein is an antibody, or a fragment thereof, labeled with a label selected from the group consisting of an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand and a radioisotope.
Claim 23
Dependent Claim
44 words
A kit for detecting a particular polynucleotide sequence in a test medium, comprising in one or more containers: (1) a nucleic acid detection probe according to claim 1, and (2) a particular protein capable of binding to the recognition site or the detection probe.
Claim 24
Dependent Claim
21 words
A kit according to claim 21, wherein the non-hybridizable portion comprises single stranded nucleic acid photochemically linked with an intercalating agent.
Claim 25
Dependent Claim
25 words
A kit according to claim 21, wherein the non-hybridizable portion comprises double stranded nucleic acid in the form of intercalating complexes with an intercalating agent.
Claim 26
Dependent Claim
26 words
A kit according to claim 23, wherein the intercalating agent is covalently linked to at least one of the two strands of the double stranded portion.
Claim 27
Dependent Claim
26 words
A kit according to claim 21, wherein the modifier compound is an intercalating agent selected from the group consisting of acridine dyes, phenazines, phenothiazines and quinolines.
Claim 28
Dependent Claim
13 words
A kit according to claim 21, wherein the particular protein carries a label.
Claim 29
Dependent Claim
33 words
A kit according to claim 26, wherein the label is selected from the group consisting of an enzymatically active group, a fluorescer, a chromophor, a luminescer, a specifically bindable ligand and a radioisotope.
Claim 30
Dependent Claim
43 words
A kit according to claim 21, wherein the protein is an antibody, or a fragment thereof, labeled with a label selected from the group consisting of an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand and a radioisotype.
Claim 31
Dependent Claim
22 words
An antibody, or fragment thereof, capable of binding with an intercalation complex formed beteween double stranded nucleic acid and a nucleic acid.
Claim 32
Dependent Claim
22 words
An antibody, or fragment thereof, capable of binding with an intercalation complex formed beteween double stranded nucleic acid and a nucleic acid.
Claim 33
Dependent Claim
18 words
The antibody or fragment thereof of claim 29 which is incapable of binding to single stranded nucleic acids.
Claim 34
Dependent Claim
24 words
An antibody or fragment thereof of claim 29 wherein the intercalator is selected from the group consisting of acridine dyes, phenazines, phenothiazines and quinolines.
Claim 35
Dependent Claim
18 words
The antibody or fragment thereof of claim 29 wherein the double stranded nucleic acid is a DNA/DNA duplex.
Claim 36
Dependent Claim
16 words
The antibody or fragment thereof of claim 29 which is labeled with a detectable chemical group.
Claim 37
Dependent Claim
37 words
An antibody or fragment thereof of claim 33 wherein the detectable chemical group is selected from the group consisting of an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand and a radioisotope.
Claim 38
Dependent Claim
85 words
A nucleic acid detection probe comprising a hybridizable single stranded protion of nucleic acid connected with a non-hybridizable, double stranded nucleic acid portion, the non-hybridizable portion including a recognition site for binding by a particular protein, wherein the non-hybridizable portion has been modified to create the protein recognition site and wherein the modification is effected by connecting to said non-hybridizable portion an intercalating agent or wherein the modification is effected by contact with a modifier compound which introduces an antigenic determinant into the non-hybridizable portion.
Claim 39
Dependent Claim
85 words
A nucleic acid detection probe comprising a hybridizable single stranded protion of nucleic acid connected with a non-hybridizable, double stranded nucleic acid portion, the non-hybridizable portion including a recognition site for binding by a particular protein, wherein the non-hybridizable portion has been modified to create the protein recognition site and wherein the modification is effected by connecting to said non-hybridizable portion an intercalating agent or wherein the modification is effected by contact with a modifier compound which introduces an antigenic determinant into the non-hybridizable portion.
Claim 40
Dependent Claim
23 words
A detection probe according to claim 35, wherein the hybridizable portion is covalently linked with one of the strands of the non-hybridizable portion.
Claim 41
Dependent Claim
22 words
A detection probes according to claim 35, wherein the hybridizable portion is complementary to the genomic sequence responsible for sickle cell anemia.
Claim 42
Dependent Claim
22 words
A detection probe according to claim 35, wherein the non-hybridizable portion comprises single stranded nucleic acid photochemically linked with an intercalating agent.
Claim 43
Dependent Claim
26 words
A detection probe according to claim 35, wherein the non-hybridizable portion comprises double stranded nucleic acid in the form of intercalating complexes with an intercalating agent.
Claim 44
Dependent Claim
27 words
A detection probe according to claim 39, wherein the intercalating agent is covalently linked to at least one of the two strands of the double stranded portion.
Claim 45
Dependent Claim
27 words
A detection probe according to claim 35, wherein the modifier compound is an intercalating agent selected from the group conisting of acridine dyes, phenazines, phenothiazines and quinolines.
Claim 46
Dependent Claim
20 words
A detection probe according to claim 35, including the particular protein bound to the protein-specific site of the non-hybridizable portion.
Claim 47
Dependent Claim
13 words
A detection probe according to claim 42, wherein the protein carries a label.
Claim 48
Dependent Claim
34 words
A detection probe according to claim 43, wherein the label is selected from the group consisting of an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand and a radioisotope.
Claim 49
Dependent Claim
62 words
A detection probe according to claim 35, including a labeled from of the particular protein bound to the protein-specific site of the non-hybridizable portion, wherein the labeled protein is an antibody, or a fragment thereof, labeled with a label selected from the group consisting of an enzymatically active group, a fluorescer, a chromophore, a luminescer, a specifically bindable ligand and a radioisotope.
References Cited (Backward Citations)
US4358535A
Cited
US4358535B1
Cited
US4395486A
Cited
EP0079139A1
Cited
EP0097373B1
Cited
GB2125964A
Cited
US4556643A
Cited
EP0131830A1
Cited
EP0146815A2
Cited
US4563417A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-10-11 | 1992-04-11 | 1992-10-11 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-10-11 | 1996-04-11 | 1996-10-11 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-10-11 | 2000-04-11 | 2000-10-11 | $7700.00 | Window Closed / Lapsed |