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Nucleic acid hybridization assay and detectable molecules useful in such assay

Assignee: Enzo Diagnostics Inc
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Filing: 1984-01-30
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Granted: 1987-11-17
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Expires: 2004-01-30
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Abstract

Abstract A detectable molecule of the formula A3-(-X-R1-E-Detb)m where A3 is A2 or a polymer, where A3 has at least one modifiable reactive group selected from the group consisting of amino, hydroxy, cis OH, halides, aryl, imidazoyl, carbonyl, carboxy, thiol or a residue comprising an activated carbon; -X- is selected from the group consisting of <IMAGE> <IMAGE> <IMAGE> <IMAGE> +TR OCONH -R1- is <IMAGE> or a C1-C10 branched or unbranched alkyl or aralkyl, which may be substituted by -OH; -Y- is a direct bond to -E-, or -Y- is -E-R2- where R2 is a C1-C10 branched or unbranched alkyl; Za is chlorine, bromine or iodine; E is O, NH or an acyclic divalent sulfur atom; Detb is a chemical moiety capable of being detected, preferably comprising biotin or a metal chelator of the formula: <IMAGE> or the 4-hydroxy or acyloxy derivative thereof, where R3 is C1-C4 alkyl or CH2COOM, M is the same or different and selected from the group consisting of hydrogen, a metal or non-metal cation or is C1-C10 alkyl, aryl or aralkyl; and m is an integer from 1 to the total number of modified reactive groups on A3. The detectable molecules are useful in in virto or in vivo assays or therapy.

Inventors

  • 👤 Jannis G. Stavrianopoulos

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-27 13:15
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/534; A; A61; A61K; A61K51/00; A61K51/02; A61K51/04; A61K51/0497; A61K51/08; A61K51/10; A61K51/1093; G01N33/533; G01N33/543; G01N33/551; G01N33/553

Claims Analysis

Total Claims: 29
Claim 1
Independent Claim 166 words
A detectable molecule of the formula A.sup.3 --(--X--R.sup.1 --E--Det.sup.b).sub.m where A 3 is an oligo- or polynucleotide having at least one modifiable reactive group consisting of amino, hydroxy, cis OH, halide, aryl, imidazoyl, carbonyl, carboxyl, thiol or a residue comprising an activated carbon; --X-- is selected from the group consisting of ##STR54## --R 1 -- is ##STR55## a C 1 -C 10 branched or unbranched alkyl or aralkyl, which may be substituted by --OH; --Y-- is a direct bond to --E--, or --Y-- is --E--R 2 -- where --R 2 -- is a C 1 -C 10 branched or unbranched alkyl; Z a is chlorine, bromine or iodine; --E-- is O, NH or an acyclic divalent sulfur atom; Det b is a chemical moiety capable of being detected, comprising biotin or a substituted or unsubstituted metal chelator or a compound capable of yielding a metal chelator; and m is an integer from 1 to the total number of modified reactive groups on said oligo- or polynucleotide.
Claim 2
Dependent Claim 166 words
A detectable molecule of the formula A.sup.3 --(--X--R.sup.1 --E--Det.sup.b).sub.m where A 3 is an oligo- or polynucleotide having at least one modifiable reactive group consisting of amino, hydroxy, cis OH, halide, aryl, imidazoyl, carbonyl, carboxyl, thiol or a residue comprising an activated carbon; --X-- is selected from the group consisting of ##STR54## --R 1 -- is ##STR55## a C 1 -C 10 branched or unbranched alkyl or aralkyl, which may be substituted by --OH; --Y-- is a direct bond to --E--, or --Y-- is --E--R 2 -- where --R 2 -- is a C 1 -C 10 branched or unbranched alkyl; Z a is chlorine, bromine or iodine; --E-- is O, NH or an acyclic divalent sulfur atom; Det b is a chemical moiety capable of being detected, comprising biotin or a substituted or unsubstituted metal chelator or a compound capable of yielding a metal chelator; and m is an integer from 1 to the total number of modified reactive groups on said oligo- or polynucleotide.
Claim 3
Dependent Claim 43 words
The molecule of claim 1 wherein said metal chelator has the formula: ##STR56## or the 4-hydroxy or acyloxy derivative thereof, where R 3 is C 1 -C 4 alkyl or is --CH 2 --COOM, and each M is a metal or non-metal cation.
Claim 4
Dependent Claim 33 words
The molecule of claim 1, wherein said activated carbon in said modifiable group is present on a base of said oligo- or polynucleotide, and is capable of covalent coupling to a diazonium compound.
Claim 5
Dependent Claim 13 words
The molecule of claim 3 wherein said base of said polynucleotide is guanine.
Claim 6
Dependent Claim 18 words
The molecule of claim 1 wherein said polynucleotide is selected from the group consisting of DNA or RNA.
Claim 7
Dependent Claim 36 words
The molecule of claim 1 wherein said modifiable reactive group is an amino group, and --X-- is selected from the group consisting of --NH--CO--, --NH--CNH--, --NHSO 2 --, --NH--N═N--, --NH--CO--CH 2 --NH--, and --NH--CO--CH 2 --S--.
Claim 8
Dependent Claim 16 words
The molecule of claim 6 wherein X is --NHCO--, --NH--COCH 2 --NH--, or --NH--CO--CH 2 --S--.
Claim 9
Dependent Claim 22 words
The molecule of claim 1 wherein said modifiable reactive group comprises an activated carbon, X is --N═N-- and R 1 is ##STR57##
Claim 10
Dependent Claim 19 words
The molecule of claim 8 wherein said residue comprising an activated carbon is a guanine, cytidine or inosine base.
Claim 11
Dependent Claim 10 words
The molecule of claim 9 wherein said base is guanine.
Claim 12
Dependent Claim 11 words
The molecule of claim 3 wherein --R 1 -- is ##STR58##
Claim 13
Dependent Claim 14 words
The molecule of claim 11 wherein R 2 is --CH 2 --CH 2 --.
Claim 14
Dependent Claim 11 words
The molecule of claim 8 wherein --R 1 -- is ##STR59##
Claim 15
Dependent Claim 11 words
The molecule of claim 13 wherein --R 1 -- is ##STR60##
Claim 16
Dependent Claim 16 words
The molecule of claim 6 wherein R 1 is a C 1 -C 10 alkyl group.
Claim 17
Dependent Claim 16 words
The molecule of claim 12 wherein R 1 is a C 1 -C 10 alkyl group.
Claim 18
Dependent Claim 24 words
The molecule of claim 7 wherein --X-- is --NH--CO--CH 2 NH-- or --NH--CO--CH 2 --S--, and R 1 is C 1 -C 6 alkyl.
Claim 19
Dependent Claim 14 words
The molecule of claim 17 wherein R 1 is --CH 2 --CH 2 --.
Claim 20
Dependent Claim 11 words
The molecule of claim 1 wherein said Det b is ##STR61##
Claim 21
Dependent Claim 10 words
The molecule of claim 1 wherein Det b is ##STR62##
Claim 22
Dependent Claim 10 words
The molecule of claim 2 wherein Det b is ##STR63##
Claim 23
Dependent Claim 10 words
The molecule of claim 2 wherein Det b is ##STR64##
Claim 24
Dependent Claim 11 words
The molecule of claim 2 wherein M is a radioactive metal.
Claim 25
Dependent Claim 13 words
The molecule of claim 1 wherein E is an acyclic divalent sulfur atom.
Claim 26
Dependent Claim 23 words
A detectable molecule according to claim 1, wherein X is ##STR65## and R 1 is C 1 -C 10 branched or unbranched alkyl.
Claim 27
Dependent Claim 19 words
A detectable molecule according to claim 1, wherein Det b comprises biotin or a substituted or unsubstituted metal chelator.
Claim 28
Dependent Claim 28 words
A detectable molecule according to claim 1, wherein said residue is selected from the group consisting of guanine and cytidine, X is --N═N--, and R 1 is ##STR66##
Claim 29
Dependent Claim 19 words
A nucleic acid hybridization assay comprising hybridizing to a nucleic acid the molecule of any one of claims 1-27.

References Cited (Backward Citations)

GB2019408A Cited
US4374120A Cited
EP0063879A2 Cited
US4563419A Cited
EP0117440A1 Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1990-11-16 1991-05-18 1991-11-17 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1994-11-16 1995-05-18 1995-11-17 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1998-11-16 1999-05-18 1999-11-17 $7700.00 Window Closed / Lapsed

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