US4746631A
Expired / Lapsed
US
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Immunoassay method, device, and test kit
Assignee: BTG International Inc
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Filing: 1985-05-09
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Granted: 1988-05-24
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Expires: 2005-05-09
Abstract
Abstract An immunoassay device includes one or more reaction chambers. Each reaction chamber is adapted to receive and retain a volume of test fluid in fluid communication with nonoverlapping first and second reaction surfaces. To the first reaction surface is immobilized analyte binding partner that is in turn saturated with analyte conjugate: analyte component conjugated to one or more components, termed ligand/marker, that serve ligand and marker functions as described herein. The analyte conjugate has a higher disassociation constant with reference to the immobilized analyte binding partner than does the analyte to be assayed. To the second reaction surface is immobilized ligand/marker binding partner. A test fluid sample is introduced into the reaction chamber and retained therein to permit two reactions to occur. In a first reaction between analyte and analyte binding partner at the first reaction reaction surface, analyte proportionately displaces analyte conjugate into the test fluid sample. In a second reaction the displaced analyte conjugate becomes sequestered at the second reaction surface by bonding with immobilized ligand/marker binding partner. Thereafter the marker activity of sequestered analyte analog is measured, the measured activity being a function of the analyte concentration that is referable to standards and controls. A test kit includes the immunoassay device in combination with comparative test results.
Inventors
- 👤 James A. Clagett
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-26 23:01
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54306; Y; Y10; Y10S; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/809
Claims Analysis
Total Claims: 55
Claim 1
Independent Claim
239 words
A method of detecting an analyte in a test fluid, comprising the steps of: providing a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a predetermined volume of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon; introducing a volume of the test fluid into the reaction chamber in fluid communication with the first and second reaction surfaces; retaining the volume of test fluid in the reaction chamber to permit two reactions to occur: a first reaction between analyte and analyte binding partner at the first reaction surface, the analyte thereby proportionally displacing analyte conjugate into the volume of test fluid, and a second reaction between the displaced analyte conjugate and ligand/marker binding partner at the second reaction surface, the displaced analyte conjugate thereby becoming sequestered on the second reaction surface; and, thereafter measuring the activity of sequestered ligand/marker on the second reaction surface, the measured activity being a function of the analyte concentration that is referable to standards and controls.
Claim 2
Independent Claim
239 words
A method of detecting an analyte in a test fluid, comprising the steps of: providing a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a predetermined volume of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon; introducing a volume of the test fluid into the reaction chamber in fluid communication with the first and second reaction surfaces; retaining the volume of test fluid in the reaction chamber to permit two reactions to occur: a first reaction between analyte and analyte binding partner at the first reaction surface, the analyte thereby proportionally displacing analyte conjugate into the volume of test fluid, and a second reaction between the displaced analyte conjugate and ligand/marker binding partner at the second reaction surface, the displaced analyte conjugate thereby becoming sequestered on the second reaction surface; and, thereafter measuring the activity of sequestered ligand/marker on the second reaction surface, the measured activity being a function of the analyte concentration that is referable to standards and controls.
Claim 3
Dependent Claim
13 words
The method of claim 1, wherein the analyte binding partner is an antibody.
Claim 4
Dependent Claim
15 words
The method of claim 1, wherein the analyte binding partner is an antigen or hapten.
Claim 5
Dependent Claim
13 words
The method of claim 1, wherein the reaction chamber is a test tube.
Claim 6
Dependent Claim
14 words
The method of claim 1, wherein the first and second reaction surfaces are noncontiguous.
Claim 7
Dependent Claim
11 words
The method of claim 1, wherein the analyte conjugate comprises phthalocyanine.
Claim 8
Dependent Claim
24 words
The method of claim 1, wherein the test fluid volume capacity of the reaction chamber is on the order of 10 to 50 μl.
Claim 9
Dependent Claim
25 words
The method of claim 7, wherein the first and second reaction surfaces are separated by a diffusion distance of no more than about 1.0 mm.
Claim 10
Dependent Claim
16 words
The method of claim 8, wherein the diffusion distance is on the order of 0.1 mm.
Claim 11
Dependent Claim
22 words
The method of claim 7, wherein the reaction chamber further comprises wick means for receiving and retaining the volume of test fluid.
Claim 12
Dependent Claim
28 words
The method of claim 10, wherein the reaction chamber further comprises valve means for releasing from the reaction chamber air displaced by the received volume of test fluid.
Claim 13
Dependent Claim
16 words
The method of claim 7, wherein the reaction chamber is detachably mounted on an insoluble support.
Claim 14
Dependent Claim
16 words
The method of claim 12, wherein the second reaction surface is affixed to the insoluble support.
Claim 15
Dependent Claim
18 words
The method of claim 13 wherein the insoluble support further comprises means for storing substrate reactive with ligand/marker.
Claim 16
Independent Claim
111 words
A device for detecting an analyte in a test fluid, comprising a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a predetermined volume of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon.
Claim 17
Independent Claim
111 words
A device for detecting an analyte in a test fluid, comprising a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a predetermined volume of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon.
Claim 18
Dependent Claim
13 words
The device of claim 15, wherein the analyte binding partner is an antibody.
Claim 19
Dependent Claim
15 words
The device of claim 15, wherein the analyte binding partner is an antigen or hapten.
Claim 20
Dependent Claim
13 words
The device of claim 15, wherein the reaction chamber is a test tube.
Claim 21
Dependent Claim
11 words
The device of claim 15, wherein the analyte conjugate comprises phthalocyanine.
Claim 22
Dependent Claim
24 words
The device of claim 15, wherein the test fluid volume capacity of the reaction chamber is on the order of 10 to 50 μl.
Claim 23
Dependent Claim
25 words
The device of claim 20, wherein the first and second reaction surfaces are separated by a diffusion distance of no more than about 1.0 mm.
Claim 24
Dependent Claim
16 words
The device of claim 21, wherein the diffusion distance is on the order of 0.1 mm.
Claim 25
Dependent Claim
18 words
The device of claim 20 further comprising wick means for receiving and retaining the volume of test fluid.
Claim 26
Dependent Claim
24 words
The device of claim 23 further comprising valve means for releasing from the reaction chamber air displaced by the received volume of test fluid.
Claim 27
Dependent Claim
16 words
The device of claim 20, wherein the reaction chamber is detachably mounted on an insoluble support.
Claim 28
Dependent Claim
16 words
The device of claim 25, wherein the second reaction surface is affixed to the insoluble support.
Claim 29
Dependent Claim
14 words
The device of claim 26 further comprising means for storing substrate reactive with ligand/marker.
Claim 30
Independent Claim
246 words
A method of detecting an analyte in a test fluid, comprising the steps of: providing a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume on the order of 100 to 200 μl of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon; introducing a volume of the test fluid into the reaction chamber in fluid communication with the first and second reaction surfaces; retaining the volume of test fluid in the reaction chamber to permit two reactions to occur: a first reaction between analyte and analyte binding partner at the first reaction surface, the analyte thereby proportionally displacing analyte conjugate into the volume of test fluid, and a second reaction between the displaced analyte conjugate and ligand/marker binding partner at the second reaction surface, the displaced analyte conjugate thereby becoming sequestered on the second reaction surface; and, thereafter measuring the activity of sequestered ligand/marker on the second reaction surface, the measured activity being a function of the analyte concentration that is referable to standards and controls.
Claim 31
Independent Claim
246 words
A method of detecting an analyte in a test fluid, comprising the steps of: providing a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume on the order of 100 to 200 μl of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon; introducing a volume of the test fluid into the reaction chamber in fluid communication with the first and second reaction surfaces; retaining the volume of test fluid in the reaction chamber to permit two reactions to occur: a first reaction between analyte and analyte binding partner at the first reaction surface, the analyte thereby proportionally displacing analyte conjugate into the volume of test fluid, and a second reaction between the displaced analyte conjugate and ligand/marker binding partner at the second reaction surface, the displaced analyte conjugate thereby becoming sequestered on the second reaction surface; and, thereafter measuring the activity of sequestered ligand/marker on the second reaction surface, the measured activity being a function of the analyte concentration that is referable to standards and controls.
Claim 32
Dependent Claim
13 words
The method of claim 28, wherein the reaction chamber is a microtiter well.
Claim 33
Dependent Claim
11 words
The method of claim 28, wherein the analyte conjugate comprises phthalocyanine.
Claim 34
Independent Claim
255 words
The method of detecting an analyte in a test fluid, comprising the steps of: providing a reaction chamber detachably mounted on an insoluble support, the reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume on the order of 10 to 50 μl of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon; introducing a volume of the test fluid into the reaction chamber in fluid communication with the first and second reaction surfaces; retaining the volume of test fluid in the reaction chamber to permit two reactions to occur: a first reaction between analyte and analyte binding partner at the first reaction surface, the analyte thereby proportionally displacing analyte conjugate into the volume of test fluid, and a second reaction between the displaced analyte conjugate and ligand/marker binding partner at the second reaction surface, the displaced analyte conjugate thereby becoming sequestered on the second reaction surface; and, thereafter measuring the activity of sequestered ligand/marker on the second reaction surface, the measured activity being a function of the analyte concentration that is referable to standards and controls.
Claim 35
Independent Claim
255 words
The method of detecting an analyte in a test fluid, comprising the steps of: providing a reaction chamber detachably mounted on an insoluble support, the reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume on the order of 10 to 50 μl of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon; introducing a volume of the test fluid into the reaction chamber in fluid communication with the first and second reaction surfaces; retaining the volume of test fluid in the reaction chamber to permit two reactions to occur: a first reaction between analyte and analyte binding partner at the first reaction surface, the analyte thereby proportionally displacing analyte conjugate into the volume of test fluid, and a second reaction between the displaced analyte conjugate and ligand/marker binding partner at the second reaction surface, the displaced analyte conjugate thereby becoming sequestered on the second reaction surface; and, thereafter measuring the activity of sequestered ligand/marker on the second reaction surface, the measured activity being a function of the analyte concentration that is referable to standards and controls.
Claim 36
Dependent Claim
11 words
The method of claim 31, wherein the analyte conjugate comprises phthalocyanine.
Claim 37
Dependent Claim
16 words
The method of claim 31, wherein the reaction chamber is detachably mounted on an insoluble support.
Claim 38
Dependent Claim
17 words
The method of claim 33, wherein the first reaction surface is detachably mounted on an insoluble support.
Claim 39
Dependent Claim
19 words
The method of claim 33, wherein a plurality of the reaction chambers are detachably mounted on an insoluble support.
Claim 40
Dependent Claim
19 words
The method of claim 35, wherein the reaction chambers have identical ligand/marker binding partners but different analyte binding partners.
Claim 41
Independent Claim
118 words
A device for detecting an analyte in a test fluid, comprising a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume on the order of 100 to 200 μl of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon.
Claim 42
Independent Claim
118 words
A device for detecting an analyte in a test fluid, comprising a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume on the order of 100 to 200 μl of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon.
Claim 43
Dependent Claim
13 words
The device of claim 37, wherein the reaction chamber is a microtiter well.
Claim 44
Dependent Claim
11 words
The device of claim 37, wherein the analyte conjugate comprises phthalocyanine.
Claim 45
Independent Claim
127 words
The device for detecting an analyte in a test fluid, comprising a reaction chamber detachably mounted on an insoluble support, the reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume on the order of 10 to 50 μl of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon.
Claim 46
Independent Claim
127 words
The device for detecting an analyte in a test fluid, comprising a reaction chamber detachably mounted on an insoluble support, the reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume on the order of 10 to 50 μl of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon.
Claim 47
Dependent Claim
16 words
The device of claim 40, wherein the second reaction surface is affixed to the insoluble support.
Claim 48
Dependent Claim
17 words
The device of claim 40, wherein the first reaction surface is detachably mounted on the insoluble support.
Claim 49
Dependent Claim
19 words
The device of claim 40, wherein a plurality of the reaction chambers are detachably mounted on the insoluble support.
Claim 50
Dependent Claim
19 words
The device of claim 43, wherein the reaction chambers have identical ligand/marker binding partners but different analyte binding partners.
Claim 51
Dependent Claim
11 words
The device of claim 40, wherein the analyte conjugate comprises phthalocyanine.
Claim 52
Dependent Claim
134 words
A test kit comprising a device for detecting an analyte in a test fluid, the device comprising a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon, in combination with comparative test results that associate ligand/marker activity with specific concentrations of analyte in test fluid.
Claim 53
Dependent Claim
134 words
A test kit comprising a device for detecting an analyte in a test fluid, the device comprising a reaction chamber having nonoverlapping first and second reaction surfaces, the reaction chamber being adapted to receive and retain a volume of the test fluid in fluid communication with the first and second reaction surfaces, the first reaction surface having an analyte binding partner immobilized thereon, the analyte binding partner having an analyte conjugate reversibly bound thereto, the analyte conjugate comprising a ligand/marker conjugated to an analyte component, the analyte conjugate having a higher disassociation constant than the analyte with reference to the immobilized analyte binding partner, and the second reaction surface having a ligand/marker binding partner immobilized thereon, in combination with comparative test results that associate ligand/marker activity with specific concentrations of analyte in test fluid.
Claim 54
Dependent Claim
15 words
The test kit of claim 46, wherein the comparative test result comprises a color chart.
Claim 55
Dependent Claim
12 words
The test kit of claim 46, wherein the analyte conjugate comprises phthalocyanine.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1991-05-24 | 1991-11-23 | 1992-05-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1995-05-24 | 1995-11-23 | 1996-05-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1999-05-24 | 1999-11-23 | 2000-05-24 | $7700.00 | Window Closed / Lapsed |