US5232830A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Intrinsic fluorescent quenching methods
Assignee: Becton Dickinson and Co
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Filing: 1990-07-26
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Granted: 1993-08-03
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Expires: 2010-07-26
Abstract
Abstract Novel methods and compositions for detecting a member of a ligand pair on solid supports having intrinsic fluorescence are disclosed. A target member of a ligand pair is contacted with a capture member of a ligand pair, wherein the capture member is immobilized on a solid support having intrinsic fluorescence, and the contacted pair is in association with a colorimetric reporter, then the solid support is irradiated the solid support, and the resultant fluorescence is determined. Preferably, methods include hybridization assays of nucleic acid sequences on such solid supports. The extent of detecting a member of a ligand pair, preferably a nucleic acid sequence, is determined using a method or technique described throughout this document as fluorescent quenching. Methods and compositions pertaining to solid supports having their intrinsic or natural fluorescence quenched or masked are also described herein. The present invention has utility in detection assays for a member of a ligand pair. Such ligand pairs include, but are not limited to, antigens (or epitopes) and appropriate antibodies, complementary nucleic acid sequences, hormones and their receptors, enzymes and corresponding inhibitors, lectins and sugars, etc.
Inventors
- 👤 Jeffrey Van Ness
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 00:23
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; G01N33/531; G01N33/532; G01N33/533; G01N33/543; G01N33/54313; G01N33/58; G01N33/582; Y; Y10; Y10S; Y10S435/00; Y10S435/80
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
71 words
A method for detecting a member of a ligand pair comprising: contacting a target member of a ligand pair with a capture member of the ligand pair, said capture member immobilized on a solid support having intrinsic fluorescence, and said contacted pair in association with a colorimetric reporter; irradiating the solid support, said solid support attached to the ligand pair in association with the colorimetric reporter; and determining the resultant fluorescence.
Claim 2
Independent Claim
71 words
A method for detecting a member of a ligand pair comprising: contacting a target member of a ligand pair with a capture member of the ligand pair, said capture member immobilized on a solid support having intrinsic fluorescence, and said contacted pair in association with a colorimetric reporter; irradiating the solid support, said solid support attached to the ligand pair in association with the colorimetric reporter; and determining the resultant fluorescence.
Claim 3
Dependent Claim
28 words
The method according to claim 1 wherein the target member is an antibody and the capture member is an antigen, said antigen capable of binding with said antibody.
Claim 4
Dependent Claim
35 words
The method according to claim 1 wherein the target member is a first nucleic acid and the capture member is a second nucleic acid, said first nucleic acid and said second nucleic acid being complementary.
Claim 5
Independent Claim
65 words
A method for nucleic acid detection comprising: contacting a target nucleic acid and a complementary capture nucleic acid probe, said capture probe immobilized on a solid support having intrinsic fluorescence, and said contacted target nucleic acid-capture probe complex in association with a colorimetric reporter; irradiating the solid support, said solid support attached to the hybridized capture probe-target nucleic acid complex; and determining the resultant fluorescence.
Claim 6
Independent Claim
65 words
A method for nucleic acid detection comprising: contacting a target nucleic acid and a complementary capture nucleic acid probe, said capture probe immobilized on a solid support having intrinsic fluorescence, and said contacted target nucleic acid-capture probe complex in association with a colorimetric reporter; irradiating the solid support, said solid support attached to the hybridized capture probe-target nucleic acid complex; and determining the resultant fluorescence.
Claim 7
Dependent Claim
26 words
The method of claim 4 wherein the colorimetric reporter is attached to a signal nucleic acid probe, said signal probe complementary to the target nucleic acid.
Claim 8
Dependent Claim
36 words
The method of claim 5 wherein the solid support is selected from the group consisting of nylons, polyethylene, polyvinylchloride, polystyrene, polypropylene, acrylonitrile butadiene styrene, acrylics, acetals, polycarbonates, fluoroplastics, polyesters, phenolics, amino resins, epoxies, polyurethane, and silicones.
Claim 9
Dependent Claim
13 words
The method of claim 6 wherein the solid support is a nylon bead.
Claim 10
Dependent Claim
12 words
The method of claim 7 wherein the colorimetric reporter is an enzyme.
Claim 11
Dependent Claim
22 words
The method of claim 8 wherein the reporter enzyme is selected from the group consisting of horseradish peroxidase, alkaline phosphatase, and β-galactosidase.
Claim 12
Independent Claim
80 words
A method for detecting a member of a ligand pair comprising: immobilizing a capture member of a ligand pair on a composition comprising a solid support having its intrinsic fluorescence quenched; contacting the target member of a ligand pair with the immobilized capture member of the ligand pair, said contacted pair in association with a fluorescent reporter; irradiating the solid support, said solid support attached to the ligand pair in association with the fluorescent reporter; and determining the resultant fluorescence.
Claim 13
Independent Claim
80 words
A method for detecting a member of a ligand pair comprising: immobilizing a capture member of a ligand pair on a composition comprising a solid support having its intrinsic fluorescence quenched; contacting the target member of a ligand pair with the immobilized capture member of the ligand pair, said contacted pair in association with a fluorescent reporter; irradiating the solid support, said solid support attached to the ligand pair in association with the fluorescent reporter; and determining the resultant fluorescence.
Claim 14
Dependent Claim
35 words
The method according to claim 10 wherein the target member is a first nucleic acid and the capture member is a second nucleic acid, said first nucleic acid and said second nucleic acid being complementary.
Claim 15
Dependent Claim
14 words
The method according to claim 11 wherein resultant fluorescence is determined with a fluorometer.
Claim 16
Independent Claim
86 words
A method for quantitating the amount of fluorescent quenching product deposited on a solid support having intrinsic fluorescence, comprising: (a) depositing a colorimetric reporter as a quenching product on a solid support having intrinsic fluorescence; (b) irradiating the solid support having the colorimetric reporter with a wavelength of light that produces fluorescence of the solid support; and then (c) comparing fluorescence of the solid support having the colorimetric reporter with fluorescence from solid support lacking the colorimetric reporter, thereby quantitating the amount of deposited colorimetric reporter.
Claim 17
Independent Claim
86 words
A method for quantitating the amount of fluorescent quenching product deposited on a solid support having intrinsic fluorescence, comprising: (a) depositing a colorimetric reporter as a quenching product on a solid support having intrinsic fluorescence; (b) irradiating the solid support having the colorimetric reporter with a wavelength of light that produces fluorescence of the solid support; and then (c) comparing fluorescence of the solid support having the colorimetric reporter with fluorescence from solid support lacking the colorimetric reporter, thereby quantitating the amount of deposited colorimetric reporter.
Claim 18
Dependent Claim
11 words
The method of claim 13, wherein the solid support is nylon.
Claim 19
Dependent Claim
20 words
The method of claim 13, wherein the solid support has a diameter of about 0.01 inch to about 0.5 inch.
Claim 20
Dependent Claim
14 words
The method of claim 13, wherein the colorimetric reporter is an insoluble enzymatic product.
Claim 21
Dependent Claim
16 words
The method of claim 13, wherein the wavelength is about 240 nm to about 750 nm.
Claim 22
Dependent Claim
34 words
The method of claim 13, further including the step of separating the solid support having the colorimetric reporter from colorimetric reporter not deposited on the solid support after step (a) and before step (b).
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1996-08-02 | 1997-02-01 | 1997-08-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-08-02 | 2001-02-01 | 2001-08-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-08-02 | 2005-02-01 | 2005-08-03 | $7700.00 | Window Closed / Lapsed |