US5270164A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for enhancing the chemiluminescence of enzyme-triggered 1,2-dioxetanes and uses therefor
Assignee: EIDP Inc
•
Filing: 1993-03-01
•
Granted: 1993-12-14
•
Expires: 2013-03-01
Abstract
Abstract A process is disclosed for the enhancement of the chemiluminescence of enzyme-triggered 1,2-dioxetanes detectable within a sample. The process comprises first providing a solid support having sample disposed thereon and suitably treated with a solution including enzyme-triggered 1,2-dioxetanes. The solid support is next dried and optionally heated either simultaneously with drying or thereafter. The steps of drying and/or heating are conducted either prior to or simultaneously with detection.
Inventors
- 👤 Lynne E. Anderson
- 👤 Robert K. Kobos
- 👤 Shay E. Polsky
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 10:46
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/535; G01N33/533; Y; Y10; Y10S; Y10S435/00; Y10S435/968
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
108 words
A process for the enhancement of the chemiluminescent signal intensity of an enzyme-triggerable 1,2-dioxetane, wherein the 1,2-dioxethane are a substrate for said enzyme, comprising: (a) providing a solid hydrophobic support having enzyme disposed thereon and treating the support with a solution including 1,2-dioxetanes to enzyme-trigger the 1,2-dioxetanes to produce a semi-stable intermediate decomposable to a chemiluminescent product adsorbed on said support; (b) drying said solid support and said intermediate thereon; (c) optionally heating said solid support and said intermediate thereon simultaneously with drying or thereafter; and (d) detecting the enhanced chemiluminescence; and further that drying with or without heating is conducted either prior to or simultaneously with detection.
Claim 2
Dependent Claim
108 words
A process for the enhancement of the chemiluminescent signal intensity of an enzyme-triggerable 1,2-dioxetane, wherein the 1,2-dioxethane are a substrate for said enzyme, comprising: (a) providing a solid hydrophobic support having enzyme disposed thereon and treating the support with a solution including 1,2-dioxetanes to enzyme-trigger the 1,2-dioxetanes to produce a semi-stable intermediate decomposable to a chemiluminescent product adsorbed on said support; (b) drying said solid support and said intermediate thereon; (c) optionally heating said solid support and said intermediate thereon simultaneously with drying or thereafter; and (d) detecting the enhanced chemiluminescence; and further that drying with or without heating is conducted either prior to or simultaneously with detection.
Claim 3
Dependent Claim
13 words
The process of claim 1 wherein drying alone is conducted prior to detection.
Claim 4
Dependent Claim
20 words
The process of claim 1 wherein drying alone is conducted prior to detection and heating is conducted simultaneously with detection.
Claim 5
Dependent Claim
14 words
The process of claim 1 wherein drying and heating are conducted prior to detection.
Claim 6
Dependent Claim
13 words
The process of claim 1 wherein drying alone is conducted simultaneously with detection.
Claim 7
Dependent Claim
14 words
The process of claim 1 wherein drying and heating are conducted simultaneously with detection.
Claim 8
Dependent Claim
46 words
The process of claim 1 wherein drying is accomplished by subjecting the solid support for a suitable time and temperature to a procedure selected from the group consisting of vacuum drying, convection drying, air drying, the use of microwave energy, and the use of electrical energy.
Claim 9
Dependent Claim
18 words
The process of claim 7 wherein vacuum drying is applied for 15 to 30 minutes at ambient temperature.
Claim 10
Dependent Claim
20 words
The process of claim 7 wherein convection drying is applied in a convection oven for 5-30 minutes at 30°-100° C.
Claim 11
Dependent Claim
18 words
The process of claim 7 wherein air drying is conducted for at least 30 minutes at ambient temperature.
Claim 12
Dependent Claim
48 words
The process of claim 1 wherein heating is accomplished by introducing the solid support for a time and temperature suitable to enhance chemiluminescence to a selected level of emission to a procedure selected from the group consisting of electrical heating, infrared radiation heating, convective heating, and microwave heating.
Claim 13
Dependent Claim
15 words
The process of claim 11 wherein heating is conducted for 1-10 minutes at 40°-100° C.
Claim 14
Independent Claim
213 words
A method for the detection of a target molecule in a solid-phase assay utilizing a chemiluminescent enzyme-triggerable 1,2-dioxetane comprising: (a) providing a solid support having one member of a specific binding pair disposed thereon wherein the member of the specific binding pair is the target molecule or is able to capture the target molecule from the sample solution; (b) adding a solution containing the complimentary binding partner which is either directly conjugated to an enzyme label or which can subsequently bind a second binding partner which is conjugated to an enzyme label; (c) optionally incubating said solid support in a solution containing said second binding partner that is conjugated to an enzyme label; (d) incubating the solid support including the specific binding pair or pairs and the enzyme label in a solution containing an enzyme-triggerable 1,2-dioxetane wherein the 1,2-dioxetane is a substrate for said enzyme to produce a semi-stable intermediate decomposable to a chemiluminescent product adsorbed on said support; (e) drying said solid-support and said intermediate thereon; (f) optionally heating said solid support and said intermediate thereon simultaneously with drying or thereafter to enhance the intensity of the chemiluminescent signal; and (g) detecting the enhanced chemiluminescence; and further that drying with or without heating is conducted either prior to or simultaneously with detection.
Claim 15
Independent Claim
213 words
A method for the detection of a target molecule in a solid-phase assay utilizing a chemiluminescent enzyme-triggerable 1,2-dioxetane comprising: (a) providing a solid support having one member of a specific binding pair disposed thereon wherein the member of the specific binding pair is the target molecule or is able to capture the target molecule from the sample solution; (b) adding a solution containing the complimentary binding partner which is either directly conjugated to an enzyme label or which can subsequently bind a second binding partner which is conjugated to an enzyme label; (c) optionally incubating said solid support in a solution containing said second binding partner that is conjugated to an enzyme label; (d) incubating the solid support including the specific binding pair or pairs and the enzyme label in a solution containing an enzyme-triggerable 1,2-dioxetane wherein the 1,2-dioxetane is a substrate for said enzyme to produce a semi-stable intermediate decomposable to a chemiluminescent product adsorbed on said support; (e) drying said solid-support and said intermediate thereon; (f) optionally heating said solid support and said intermediate thereon simultaneously with drying or thereafter to enhance the intensity of the chemiluminescent signal; and (g) detecting the enhanced chemiluminescence; and further that drying with or without heating is conducted either prior to or simultaneously with detection.
Claim 16
Dependent Claim
25 words
The process of claim 13 wherein said solid support is hydrophobic and selected from the group consisting of nylon membranes, polymer beads, and microtiter plates.
Claim 17
Dependent Claim
28 words
The process of claim 13 wherein said specific binding pairs are selected from the group consisting of complimentary nucleic acid strands, antibody-antigen, binding protein-vitamin, and binding protein-nucleic acid.
Claim 18
Dependent Claim
35 words
The process of claim 13 wherein said 1,2-dioxetane is 3-(2'-spiroadamantane)-4-methoxy-4-(3"-phosphorlyoxy) phenyl-1,2-dioxetane and said second binding partner is an alkaline phosphatase conjugate, or said 1,2-dioxetane is 3-(2'-spriroadamantane)-4-methoxy-4-(3"-β-D-galactopyranosyloxy-phenyl)-1,2-dioxetane and said second binding partner is a β-D-galactosidase conjugate.
Claim 19
Dependent Claim
32 words
The process of claim 13 wherein the chemiluminescent signal intensity is detected using equipment selected from the group consisting of X-ray or photographic film, a photomultiplier tube, or an electronic imaging detector.
Claim 20
Dependent Claim
28 words
The process of claim 13 wherein the assay is selected from the group consisting of ELISA (enzyme-Linked Immunosorbent Assay), DNA probe assay, and Southern, Northern and Western analyses.
References Cited (Backward Citations)
US4705847A
Cited
WO1986003840A1
Cited
US4857652A
Cited
US4959182A
Cited
WO1988000695A1
Cited
EP0261719A2
Cited
US5089423A
Cited
WO1989006650A1
Cited
US4956477A
Cited
US4952707A
Cited
US4948975A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1996-12-13 | 1997-06-14 | 1997-12-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2000-12-13 | 2001-06-14 | 2001-12-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2004-12-13 | 2005-06-14 | 2005-12-14 | $7700.00 | Window Closed / Lapsed |