US4582792A
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US
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Immunoassay method using two immobilized biologically active substances
Assignee: Fujirebio Inc
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Filing: 1983-05-05
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Granted: 1986-04-15
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Expires: 2003-05-05
Abstract
Abstract A biologically active composition comprising an immobilized phase comprising an antigen or an antibody, and an immobilized phase comprising an enzyme, an enzyme inhibitor or activator, and a method of measuring antigen or antibody using the same. According to the method of the invention, antigen with antibody can be detected in a simple procedure and high sensitivity.
Inventors
- 👤 Yasushi Kasahara
- 👤 Hiromasa Suzuki
- 👤 Yoshihiro Ashihara
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 12:15
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/532; G01N33/535; G01N33/536; G01N33/542; Y; Y10; Y10S; Y10S436/00; Y10S436/807; Y10S436/81
Claims Analysis
Total Claims: 22
Claim 1
Independent Claim
245 words
A method of measuring the amount of an antigen which comprises the steps of: (a) forming a biologically active composition by: (i) immobilizing on a first solid phase an antibody capable of reacting with the antigen to be measured, (ii) immobilizing on a second solid phase a member selected from the group consisting of enzymes, enzyme inhibitors and enzyme activators; and wherein said first and second immobilized solid phases are separated from each other, (b) contacting said biologically active composition with an aqueous solution of a water soluble combination material and a solution of the antigen to be measured, said water soluble combination material comprised of an antigen capable of reacting with the first immobilized phase antibody, coupled with a member capable of reacting with the second immobilized phase selected from the group consisting of enzyme inhibitors, enzyme activators and enzymes with the proviso (a) that when the member immobilized on the second phase in (ii) is an enzyme, then the member of the water soluble combination material capable of reacting with the second immobilized phase is either an enzyme activator or an enzyme inhibitor and (b) that when the member immobilized on the second phase in (ii) is an enzyme activator or an enzyme inhibitor, then the member of the water soluble combination material capable of reacting with the second immobilized phase is an enzyme; and (c) measuring enzyme activity, enzyme inhibiting activity, or enzyme activating activity of the biologically active composition or solution.
Claim 2
Independent Claim
245 words
A method of measuring the amount of an antigen which comprises the steps of: (a) forming a biologically active composition by: (i) immobilizing on a first solid phase an antibody capable of reacting with the antigen to be measured, (ii) immobilizing on a second solid phase a member selected from the group consisting of enzymes, enzyme inhibitors and enzyme activators; and wherein said first and second immobilized solid phases are separated from each other, (b) contacting said biologically active composition with an aqueous solution of a water soluble combination material and a solution of the antigen to be measured, said water soluble combination material comprised of an antigen capable of reacting with the first immobilized phase antibody, coupled with a member capable of reacting with the second immobilized phase selected from the group consisting of enzyme inhibitors, enzyme activators and enzymes with the proviso (a) that when the member immobilized on the second phase in (ii) is an enzyme, then the member of the water soluble combination material capable of reacting with the second immobilized phase is either an enzyme activator or an enzyme inhibitor and (b) that when the member immobilized on the second phase in (ii) is an enzyme activator or an enzyme inhibitor, then the member of the water soluble combination material capable of reacting with the second immobilized phase is an enzyme; and (c) measuring enzyme activity, enzyme inhibiting activity, or enzyme activating activity of the biologically active composition or solution.
Claim 3
Independent Claim
261 words
A method of measuring the amount of an antigen which comprises the steps of: (a) forming a biologically active composition by: (i) immobilizing on a first solid phase an antigen capable of reacting with an antibody contained in an aqueous solution of a water soluble combination material, (ii) immobilizing on a second solid phase a member selected from the group consisting of enzymes, enzyme inhibitors and enzyme activators; and wherein said first and second immobilized solid phases are separated from each other, (b) contacting said biologically active composition with the aqueous solution of the water soluble combination material and a solution of the antigen to be measured, said water soluble combination material comprised of an antibody capable of reacting with the antigen of the first immobilized phase and the antigen to be measured, coupled with a member capable of reacting with the second immobilized phase selected from the group consisting of enzyme inhibitors, enzyme activators and enzymes with the proviso (a) that when the member immobilized on the second phase in (ii) is an enzyme, then the member of the water soluble combination material capable of reacting with the second immobilized phase is either an enzyme activator or an enzyme inhibitor and (b) that when the member immobilized on the second phase in (ii) is an enzyme activator or an enzyme inhibitor, then the member of the water soluble combination material capable of reacting with the second immobilized phase is an enzyme; and (c) measuring enzyme activity, enzyme inhibiting activity, or enzyme activating activity of the biologically active composition or solution.
Claim 4
Independent Claim
261 words
A method of measuring the amount of an antigen which comprises the steps of: (a) forming a biologically active composition by: (i) immobilizing on a first solid phase an antigen capable of reacting with an antibody contained in an aqueous solution of a water soluble combination material, (ii) immobilizing on a second solid phase a member selected from the group consisting of enzymes, enzyme inhibitors and enzyme activators; and wherein said first and second immobilized solid phases are separated from each other, (b) contacting said biologically active composition with the aqueous solution of the water soluble combination material and a solution of the antigen to be measured, said water soluble combination material comprised of an antibody capable of reacting with the antigen of the first immobilized phase and the antigen to be measured, coupled with a member capable of reacting with the second immobilized phase selected from the group consisting of enzyme inhibitors, enzyme activators and enzymes with the proviso (a) that when the member immobilized on the second phase in (ii) is an enzyme, then the member of the water soluble combination material capable of reacting with the second immobilized phase is either an enzyme activator or an enzyme inhibitor and (b) that when the member immobilized on the second phase in (ii) is an enzyme activator or an enzyme inhibitor, then the member of the water soluble combination material capable of reacting with the second immobilized phase is an enzyme; and (c) measuring enzyme activity, enzyme inhibiting activity, or enzyme activating activity of the biologically active composition or solution.
Claim 5
Dependent Claim
23 words
The method of claim 2 wherein the antigen to be measured and the antigen capable of reacting with the antibody are the same.
Claim 6
Dependent Claim
16 words
The method of claim 3 wherein the antigen is obtained from an organ, blood or urine.
Claim 7
Independent Claim
245 words
A method of measuring the amount of an antibody which comprises the steps of: (a) forming a biologically active composition by: (i) immobilizing on a first solid phase an antigen capable of reacting with the antibody to be measured, (ii) immobilizing on a second solid phase a member selected from the group consisting of enzymes, enzyme inhibitors and enzyme activators; and wherein said first and second immobilized solid phases are separated from each other, (b) contacting said biologically active composition with an aqueous solution of a water soluble combination material and a solution of the antibody to be measured, said water soluble combination material comprised of an antibody capable of reacting with the first immobilized phase antigen, coupled with a member capable of reacting with the second immobilized phase selected from the group consisting of enzyme inhibitors, enzyme activators and enzymes with the proviso (a) that when the member immobilized on the second phase in (ii) is an enzyme, then the member of the water soluble combination material capable of reacting with the second immobilized phase is either an enzyme activator or an enzyme inhibitor and (b) that when the member immobilized on the secod phase in (ii) is an enzyme activator or an enzyme inhibitor, then the member of the water soluble combination material capable of reacting with the second immobilized phase is an enzyme; and (c) measuring enzyme activity, enzyme inhibiting activity, or enzyme activating activity of the biologically active composition or solution.
Claim 8
Independent Claim
245 words
A method of measuring the amount of an antibody which comprises the steps of: (a) forming a biologically active composition by: (i) immobilizing on a first solid phase an antigen capable of reacting with the antibody to be measured, (ii) immobilizing on a second solid phase a member selected from the group consisting of enzymes, enzyme inhibitors and enzyme activators; and wherein said first and second immobilized solid phases are separated from each other, (b) contacting said biologically active composition with an aqueous solution of a water soluble combination material and a solution of the antibody to be measured, said water soluble combination material comprised of an antibody capable of reacting with the first immobilized phase antigen, coupled with a member capable of reacting with the second immobilized phase selected from the group consisting of enzyme inhibitors, enzyme activators and enzymes with the proviso (a) that when the member immobilized on the second phase in (ii) is an enzyme, then the member of the water soluble combination material capable of reacting with the second immobilized phase is either an enzyme activator or an enzyme inhibitor and (b) that when the member immobilized on the secod phase in (ii) is an enzyme activator or an enzyme inhibitor, then the member of the water soluble combination material capable of reacting with the second immobilized phase is an enzyme; and (c) measuring enzyme activity, enzyme inhibiting activity, or enzyme activating activity of the biologically active composition or solution.
Claim 9
Independent Claim
261 words
A method of measuring the amount of an antibody which comprises the steps of: (a) forming a biologically active composition by: (i) immobilizing on a first solid phase an antibody capable of reacting with an antigen contained in an aqueous solution of a water soluble combination material, (ii) immobilizing on a second solid phase a member selected from the group consisting of enzymes, enzyme inhibitors and enzyme activators; and wherein said first and second immobilized solid phases are separated from each other, (b) contacting said biologically active composition with the aqueous solution of the water soluble combination material and a solution of the antibody to be measured, said water soluble combination material comprised of an antigen capable of reacting with the antibody of the first immobilized phase and the antibody to be measured, coupled with a member capable of reacting with the second immobilized phase selected from the group consisting of enzyme inhibitors, enzyme activators and enzymes with the proviso (a) that when the member immobilized on the second phase in (ii) is an enzyme, then the member of the water soluble combination material capable of reacting with the second immobilized phase is either an enzyme activator or an enzyme inhibitor and (b) that when the member immobilized on the second phase in (ii) is an enzyme activator or an enzyme inhibitor, then the member of the water soluble combination material capable of reacting with the second immobilized phase is an enzyme; and (c) measuring enzyme activity, enzyme inhibiting activity, or enzyme activating activity of the biologically active composition or solution.
Claim 10
Independent Claim
261 words
A method of measuring the amount of an antibody which comprises the steps of: (a) forming a biologically active composition by: (i) immobilizing on a first solid phase an antibody capable of reacting with an antigen contained in an aqueous solution of a water soluble combination material, (ii) immobilizing on a second solid phase a member selected from the group consisting of enzymes, enzyme inhibitors and enzyme activators; and wherein said first and second immobilized solid phases are separated from each other, (b) contacting said biologically active composition with the aqueous solution of the water soluble combination material and a solution of the antibody to be measured, said water soluble combination material comprised of an antigen capable of reacting with the antibody of the first immobilized phase and the antibody to be measured, coupled with a member capable of reacting with the second immobilized phase selected from the group consisting of enzyme inhibitors, enzyme activators and enzymes with the proviso (a) that when the member immobilized on the second phase in (ii) is an enzyme, then the member of the water soluble combination material capable of reacting with the second immobilized phase is either an enzyme activator or an enzyme inhibitor and (b) that when the member immobilized on the second phase in (ii) is an enzyme activator or an enzyme inhibitor, then the member of the water soluble combination material capable of reacting with the second immobilized phase is an enzyme; and (c) measuring enzyme activity, enzyme inhibiting activity, or enzyme activating activity of the biologically active composition or solution.
Claim 11
Dependent Claim
64 words
The method of claim 2, 5 or 6 wherein the second immobilized solid phase comprises an enzyme or enyzme inhibitor and said water soluble combination comprises an enzyme inhibitor or enzyme capable of reacting with the second immobilized solid phase coupled with the antigen or antibody and the enzyme activity or enyzme inhibiting activity of the biologically active composition or the solution is measured.
Claim 12
Dependent Claim
11 words
The method of claim 7 wherein the enzyme activity is measured.
Claim 13
Dependent Claim
31 words
The method of claim 7 wherein the first immobilized phase antibody or water soluble combination material antibody is F(ab') 2 , Fab', Fab or a second antibody against a first antibody.
Claim 14
Dependent Claim
13 words
The method of claim 7 wherein the enzyme is α-amylase, β-galactosidase or esterase.
Claim 15
Dependent Claim
36 words
The method of claim 7 wherein the enzyme is selected from the groups consisting of α-amylase, β-galactosidase and esterase and wherein the enzyme inhibitor is selected from the group consisting of α-amylase inhibitor, isoflavonoid and bestatin.
Claim 16
Dependent Claim
35 words
The method of claim 7 wherein the biologically active composition is a polymer on which the antigen or antibody and the enzyme or enzyme inhibitor are separately immobilized on parts of the same polymer particle.
Claim 17
Dependent Claim
14 words
The method of claim 12 wherein the polymer is a block-copolymer or a graft-copolymer.
Claim 18
Dependent Claim
43 words
The method of claim 7 wherein the biologically active composition comprises two polymer particles adhering to each other, on one of the polymer particles the antigen or antibody is immobilized and on the other polymer particle, the enzyme or enzyme inhibitor is immobilized.
Claim 19
Dependent Claim
37 words
The method of claim 7 wherein the biologically active composition comprises two separate particles on one of the particles the antigen or antibody is immobilized and on the other particle the enzyme or enzyme inhibitor is immobilized.
Claim 20
Dependent Claim
21 words
The method of claim 1 or 2 wherein the antigen to be measured is obtained from an organ, blood or urine.
Claim 21
Dependent Claim
41 words
The method of claim 16 wherein the antigen to be measured is a hormone derived from an endocrine gland, a blood serum protein, an immune complex, a pathogen, a complement, a drug, a cyclic nucleotide, an α-fetoprotein or a carcinoembryonic antigen.
Claim 22
Dependent Claim
22 words
The method of claim 17 wherein the antigen to be measured is ferritin, β 2 -microglobulin, α-fetoprotein, human IgG or human IgE.
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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) | 1989-04-14 | 1989-10-14 | 1990-04-15 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-04-14 | 1993-10-14 | 1994-04-15 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-04-14 | 1997-10-14 | 1998-04-15 | $7700.00 | Window Closed / Lapsed |