EP0455905A2
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EP
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Solid supports for nucleic acid hybridization assays and methods for covalently immobilizing oligonucleotides
Assignee: Epoch Biosciences Inc
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Filing: 1990-09-04
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Granted: —
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Expires: 2010-09-04
Abstract
Abstract Compositions and methods for covalently immobilizing an oligonucleotide onto a polymer-coated bead or similar structure are described. Specifically, the polymer-coated bead or similar structure possesses a large number of activatable moieties, preferably primary and secondary amines. An oligonucleotide is activated with a monofunctional or multi functional reagent, preferably the homotrifunctional reagent cyanuric chloride. Additionally, dichlorotriazine oligonucleotides and processes for activating oligonucleotides by treatment with cyanuric chloride are included in the present invention.
Inventors
- 👤 Jeffrey Van Ness
- 👤 John C. Tabone
- 👤 Charles E. Petrie
- 👤 Nicolaas M.J. Vermeulen
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-06 02:33
CPC Classifications
C; C07; C07H; C07H21/00; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6834; C12Q1/6876; C12Q1/6888; C12Q1/689; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54313; G01N33/58; G01N33/582
Claims Analysis
Total Claims: 41
Claim 1
Independent Claim
11 words
A composition comprising an oligonucleotide covalently attached to a polymer-coated bead.
Claim 2
Dependent Claim
17 words
The composition according to claim 1 wherein the oligonucleotide is activated with a monofunctional or multifunctional reagent.
Claim 3
Dependent Claim
11 words
The composition according to claim 2 wherein the reagent is homotrifunctional.
Claim 4
Dependent Claim
12 words
The composition according to claim 3 wherein the reagent is cyanuric chloride.
Claim 5
Dependent Claim
13 words
The composition acording to claim 1 wherein the polymer is an amine-containing polymer.
Claim 6
Dependent Claim
14 words
The composition according to claim 5 wherein the polymer is activated with cyanuric chloride.
Claim 7
Dependent Claim
16 words
The composition according to claim 6 wherein the polymer-coated bead is nylon and spherical in shape.
Claim 8
Dependent Claim
24 words
The composition according to claim 7 wherein the polymer-coated bead has a diameter in the range from about 0.01 inch to about 0.5 inch.
Claim 9
Dependent Claim
21 words
The composition according to claim 8 wherein the polymer-coated bead has a diameter from about 0.06 inch to about 0.09 inch.
Claim 10
Dependent Claim
17 words
The composition according to claim 9 wherein the polymer-coated bead has a diameter of about 0.09 inch.
Claim 11
Dependent Claim
17 words
The composition according to claim 10 wherein the oligonucleotide is activated through a 5'- or 3'-tethered amine.
Claim 12
Dependent Claim
16 words
The composition according to claim 10 wherein the oligonucleotide is activated through an internal tethered amine.
Claim 13
Dependent Claim
15 words
The composition according to claim 10 wherein the covalent attachment is through a spacer arm.
Claim 14
Dependent Claim
20 words
A dipstick for a nucleic acid hybridization assay comprising a nonporous solid support and means for attaching a polymer-coated bead.
Claim 15
Dependent Claim
18 words
A dipstick for a nucleic acid hybridization assay comprising a nonporous solid support having at least one depression.
Claim 16
Dependent Claim
18 words
A dipstick for a nucleic acid hybridization assay comprising a nonporous solid support having at least one perforation.
Claim 17
Dependent Claim
15 words
The dipstick according to claim 16 further comprising a polymer-coated bead attached to the perforation.
Claim 18
Dependent Claim
16 words
The dipstick according to claim 17 wherein the polymer-coated bead is covalently attached to an oligonucleotide.
Claim 19
Dependent Claim
26 words
The dipstick according to claim 18 wherein the polymer-coated bead is covalently attached to more than one oligonucleotide having the same or different nucleic acid sequences.
Claim 20
Dependent Claim
16 words
The dipstick according to claim 19 wherein the polymer-coated bead is nylon and spherical in shape.
Claim 21
Dependent Claim
25 words
The dipstick according to claim 20 wherein the diameter of the polymer-coated bead is in the range from about 0.01 inch to about 0.5 inch.
Claim 22
Dependent Claim
21 words
The dipstick according to claim 21 wherein the diameter of the bead is from about 0.06 inch to about 0.09 inch.
Claim 23
Dependent Claim
24 words
The dipstick according to claim 22 wherein there is more than one perforation and each perforation has a polymer-coated bead attached to the perforation.
Claim 24
Dependent Claim
16 words
The dipstick according to claim 23 wherein there are from about two to about ten perforations.
Claim 25
Dependent Claim
18 words
The dipstick according to claim 24 wherein the perforations are linearly situated along one edge of the dipstick.
Claim 26
Dependent Claim
19 words
The dipstick according to claim 25 wherein the polymer-coated beads are covalently attached through a spacer arm to oligonucleotides.
Claim 27
Dependent Claim
30 words
The dipstick according to claim 26 wherein at least one polymer-coated bead is covalently attached to an oligonucleotide having at least one different nucleic acid sequence from the other oligonucleotides.
Claim 28
Dependent Claim
27 words
The dipstick according to claim 27 wherein the nucleic acid sequences of the activated oligonucleotides are selected from organisms or cell types having pathogenic or nonpathogenic characteristics.
Claim 29
Dependent Claim
6 words
A composition comprising a dichlorotriazine oligonucleotide.
Claim 30
Dependent Claim
73 words
A process for covalently immobilizing an oligonucleotide on a nylon solid support comprising: treating the nylon solid support with an alkylating agent; reacting the treated solid support with an amine-containing polymer, whereby the polymer coats the solid support; activating an oligonucleotide with a homofunctional or heterofunctional reagent; conjugating the activated oligonucleotide and the polymer-coated solid support; and adding a blocking agent to the polymer-coated bead after conjugation, whereby unreacted amines are rendered nonreactive.
Claim 31
Dependent Claim
13 words
The process according to claim 30 wherein the solid support is a bead.
Claim 32
Dependent Claim
13 words
The process according to claim 31 wherein the alkylating agent is trialkyloxonium salt.
Claim 33
Dependent Claim
20 words
The process according to claim 32 wherein the polymer is selected from the group consisting of poly(ethyleneimine), polyvinylamine, and polyallylamine.
Claim 34
Dependent Claim
11 words
The process according to claim 33 wherein the reagent is homotrifunctional.
Claim 35
Dependent Claim
12 words
The process according to claim 34 wherein the reagent is cyanuric chloride.
Claim 36
Dependent Claim
13 words
The process according to claim 35 wherein the blocking agent is succinic anhydride.
Claim 37
Dependent Claim
28 words
A process for nucleic acid detection comprising: contacting a composition of claim 1 with a target nucleic acid under suitable conditions for hybridization; and detecting the hybridized product.
Claim 38
Dependent Claim
14 words
The process according to claim 37 wherein the process occurs in a microtiter well.
Claim 39
Dependent Claim
14 words
The process according to claim 37 wherein the process occurs in a flow-through tube.
Claim 40
Dependent Claim
26 words
The process according to claim 37 wherein the composition comprising an oligonucleotide covalently attached to a polymer-coated bead is attached to a perforation in a dipstick.
Claim 41
Dependent Claim
13 words
A process for activating an oligonucleotide comprising treating the oligonucleotide with cyanuric chloride.
References Cited (Backward Citations)
No citation records indexed.
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