EP0302175A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Modified labeled nucleotides and polynucleotides and methods of preparing, utilizing and detecting same
Assignee: Enzo Biochem Inc
•
Filing: 1983-06-22
•
Granted: —
•
Expires: 2003-06-22
Abstract
Abstract Nucleotides and polynucleotides, including DNA, are chemically modified or labeled so as to be capable of ready detection when attached to and/or incorporated in nucleic acid material.
Inventors
- 👤 Dean Engelhardt
- 👤 Elazar Rabbani
- 👤 Stanley Kline
- 👤 Jannis G. Stavrianopoulos
- 👤 Dollie Kirtikar
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-29 08:20
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6816; A; A61; A61K; A61K47/00; A61K47/50; A61K47/51; A61K47/62; C07; C07B; C07B59/00; C07B59/008; C07H; C07H19/00; C07H19/02; C07H19/04; C07H19/06; C07H19/10; C07K; C07K17/00; C07K17/02; C07K17/06; C12Q1/6804
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
230 words
A nucleotide having the formula P-S-B-Sig wherein P is a phosphoric acid moiety, S is a sugar moiety, B is a pyrimidine, purine or 7-deazapurine moiety, P being attached at the 3ʹ and/or 5ʹ position of S when said nucleotide is a deoxyribonucleotide and at the 2ʹ, 3ʹ and/or 5ʹ position when said nucleotide is a ribonucleotide, B being attached to the 1ʹ position of S from the N₁ position when B is a pyrimidine or the N9 position when B is a purine or a 7-deazapurine, and Sig is covalently attached to B at the N3, C5 or C6 position when B is a pyrimidine, at the C2, N3, N7 or C8 position when B is a purine, at the C7 position when B is a 7-deazapurine, at the N₂ position when B is a guanine and at the N6 position when B is an adenine, and wherein Sig represents a moiety which is detectable when said nucleotide is incorporated into a double-stranded nucleic acid duplex directly or through a linkage group which does not interfere substantially with the characteristic ability of Sig to form a detectable signal, said Sig comprising a component selected from the group consisting of biotin or iminobiotin, an electron dense component, a magnetic component, an enzyme component, a catalytic metal component, a fluorescent component, a chemiluminescent component and an antigen, hapten or antibody component.
Claim 2
Dependent Claim
230 words
A nucleotide having the formula P-S-B-Sig wherein P is a phosphoric acid moiety, S is a sugar moiety, B is a pyrimidine, purine or 7-deazapurine moiety, P being attached at the 3ʹ and/or 5ʹ position of S when said nucleotide is a deoxyribonucleotide and at the 2ʹ, 3ʹ and/or 5ʹ position when said nucleotide is a ribonucleotide, B being attached to the 1ʹ position of S from the N₁ position when B is a pyrimidine or the N9 position when B is a purine or a 7-deazapurine, and Sig is covalently attached to B at the N3, C5 or C6 position when B is a pyrimidine, at the C2, N3, N7 or C8 position when B is a purine, at the C7 position when B is a 7-deazapurine, at the N₂ position when B is a guanine and at the N6 position when B is an adenine, and wherein Sig represents a moiety which is detectable when said nucleotide is incorporated into a double-stranded nucleic acid duplex directly or through a linkage group which does not interfere substantially with the characteristic ability of Sig to form a detectable signal, said Sig comprising a component selected from the group consisting of biotin or iminobiotin, an electron dense component, a magnetic component, an enzyme component, a catalytic metal component, a fluorescent component, a chemiluminescent component and an antigen, hapten or antibody component.
Claim 3
Dependent Claim
14 words
An oligonucleotide or polynucleotide comprising at least one nucleotide in accordance with claim 1.
Claim 4
Dependent Claim
22 words
An oligo- or polynucleotide comprising at least one nucleotide of claim 1 wherein said oligo- or polynucleotide is attached to a polypeptide.
Claim 5
Dependent Claim
14 words
The oligo- or polynucleotide of claim 3, wherein said polypeptide is terminally attached thereto.
Claim 6
Dependent Claim
13 words
The oligo- or polynucleotide of claim 3 wherein said polypeptide comprises a polylysine.
Claim 7
Dependent Claim
24 words
The oligo- or polynucleotide of claim 3, wherein said polypeptide has at least one biotin, iminobiotin, antibiotin, antiiminobiotin, avidin, streptavidin or enzyme attached thereto.
Claim 8
Dependent Claim
31 words
Use of a nucleotide or polynucleotide according to one of the preceding claims for the manufacture of a chemotherapeutic agent for inhibiting RNA and/or DNA synthesis or function in an organism.
Claim 9
Dependent Claim
37 words
Use of a nucleotide or polynucleotide according to one of the preceding claims for the manufacture of a stimulating or inducing agent for the stimulation or induction of cells for the production of lymphokines, cytokines and/or interferon.
Claim 10
Dependent Claim
61 words
A method of detecting a first compound which includes a nucleotide in accordance with claim 1 as part of said first compound, which comprises contacting said first compound with a second compound capable of forming a complex therewith under suitable conditions so as to form said complex, said complex comprising said first compound and said second compound and detecting said complex.
Claim 11
Dependent Claim
55 words
A method of determining the presence of a deoxyribonucleic or ribonucleic acid molecule which comprises forming a double-stranded hybrid polynucleotide duplex which includes a single strand of deoxyribonucleic acid or ribonucleic acid corresponding to or derived from said deoxyribonucleic or ribonucleic acid molecule and a nucleotide in accordance with claim 1 and detecting said duplex.
Claim 12
Dependent Claim
99 words
A method of detecting the presence of a nucleic acid-containing etiological agent in a subject which comprises obtaining a suitable sample from said subject, detecting the presence in said sample of a deoxyribonucleic or ribonucleic acid naturally associated with said etiological agent by forming under suitable conditions a double-stranded polynucleotide duplex which includes a compound in accordance with claim 1 and a single strand of deoxyribonucleic or ribonucleic acid corresponding to or derived from said deoxyribonucleic or ribonucleic acid which is naturally associated with said etiological agent under suitable conditions and detecting the presence of said double-stranded polynucleotide duplex.
Claim 13
Dependent Claim
106 words
A method of testing a bacterium to determine its resistance to an antibiotic which comprises preparing a polynucleotide complementary to the deoxyribonucleic acid gene sequence of said bacterium which confers resistance of said bacterium to said antibiotic and which includes a nucleotide in accordance with claim 1 incorporated therein, contacting said polynucleotide under suitable conditions with a deoxyribonucleic acid obtained from said bacterium so as to form a double-stranded hybrid duplex and detecting the presence of said duplex, the detection of said duplex indicating resistance of said bacterium to said antibiotic and the absence of said duplex indicating the susceptibility of said bacterium to an antibiotic.
Claim 14
Dependent Claim
89 words
A method of diagnosing a genetic disorder in a subject which comprises preparing a polynucleotide complementary to the deoxyribonucleic acid gene sequence of said subject which is associated with said genetic disorder and includes a compound in accordance with claim 1 incorporated therein, contacting said polynucleotide under suitable conditions with deoxyribonucleic acid obtained from said subject so as to form a double-stranded hybrid duplex and detecting the presence of said hybrid duplex, the presence or absence of said hydrid duplex indicating the presence or absence of said genetic disorder.
Claim 15
Dependent Claim
74 words
A method of diagnosing thalassemia in a human subject which comprises preparing a polynucleotide complementary to the deoxyribonucleic acid gene sequence which is absent in β-minus-thalassemia subjects and includes a nucleotide in accordance with claim 1, contacting said polynucleotide under suitable conditions with deoxyribonucleic acid obtained from said subject so as to form a double-stranded hybrid duplex and determining the presence of said duplex, the absence of said duplex indicating the presence of β-minus-thalassemia.
Claim 16
Dependent Claim
77 words
A method of chromosomal karyotyping which comprises preparing a series of modified polynucleotides corresponding to a series of defined genetic sequences located on chromosomes, said polynucleotides including one or more compounds in accordance with claim 1, contacting said polynucleotides with deoxyribonucleic acid of or obtained from chromosomes so as to form hybrid duplexes and detecting said duplexes, thereby determining the location of said duplexes on said chromosomes and the location of said genetic sequences on said chromosomes.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.