Patent US4098876A successfully refreshed from Google Patents.
US4098876A
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US
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Reverse sandwich immunoassay
Assignee: Bayer Corp
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Filing: 1976-10-26
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Granted: 1978-07-04
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Expires: 1996-10-26
Abstract
Abstract Method of determining the presence and/or concentration of a polyvalent antigenic substance in a fluid. The method comprises the steps of incubating the fluid with labeled antibodies to the substance to form a first labeled immunochemical complex and then incubating that complex with immobilized antibodies to the substance to form a second labeled complex which is separated from the incubation medium. The amount of label in the second complex provides a means for detecting and/or quantitating the substance in the fluid. By reversing the sequence of incubation steps in a "two-site" or sandwich assay, greater sensitivity is achieved and an intermediate washing step is eliminated.
Inventors
- 👤 Roger N. Piasio
- 👤 James W. Ryan
- 👤 James E. Woiszwillo
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 17:15
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/551; G01N33/552; G01N33/536; G01N33/537; G01N33/538; G01N33/54306; G01N33/74; G01N33/78; G01N2470/00; G01N2470/04; Y; Y10; Y10S; Y10S436/00; Y10S436/82
Claims Analysis
Total Claims: 35
Claim 1
Independent Claim
125 words
A method of determining the presence or concentration of a polyvalent antigenic substance in a fluid sample which comprises the steps of: (a) incubating the fluid sample with labeled antibodies to the substance under conditions sufficient to form a first labeled immunochemical complex; (b) incubating that complex with a composite comprising antibodies to the substance immobilized on a water insoluble carrier material, the incubation being under conditions sufficient to form a second labeled complex comprising the first complex complexed to the composite; (c) separating the second labeled complex from the incubation medium; (d) determining the amount of label associated with the second complex; (e) relating the determination of step (d) to a standard to determine the presence or concentration of the substance in the fluid.
Claim 2
Independent Claim
125 words
A method of determining the presence or concentration of a polyvalent antigenic substance in a fluid sample which comprises the steps of: (a) incubating the fluid sample with labeled antibodies to the substance under conditions sufficient to form a first labeled immunochemical complex; (b) incubating that complex with a composite comprising antibodies to the substance immobilized on a water insoluble carrier material, the incubation being under conditions sufficient to form a second labeled complex comprising the first complex complexed to the composite; (c) separating the second labeled complex from the incubation medium; (d) determining the amount of label associated with the second complex; (e) relating the determination of step (d) to a standard to determine the presence or concentration of the substance in the fluid.
Claim 3
Dependent Claim
17 words
The method of claim 1 wherein the labeled antibodies of step (a) are labeled with an enzyme.
Claim 4
Dependent Claim
18 words
The method of claim 1 wherein the labeled antibodies of step (a) are labeled with a fluorogenic material.
Claim 5
Dependent Claim
17 words
The method of claim 1 wherein the labeled antibodies of step (a) are labeled with a radioisotope.
Claim 6
Dependent Claim
18 words
The method of claim 1 wherein the composite of step (b) comprises antibodies immobilized on an organic carrier.
Claim 7
Dependent Claim
18 words
The method of claim 1 wherein the composite of step (b) comprises antibodies immobilized on an inorganic carrier.
Claim 8
Dependent Claim
11 words
The method of claim 6 wherein the carrier comprises glass particles.
Claim 9
Dependent Claim
14 words
The method of claim 1 wherein the determination of step (d) is done enzymatically.
Claim 10
Dependent Claim
14 words
The method of claim 1 wherein the determination of step (d) is done fluorometrically.
Claim 11
Dependent Claim
14 words
The method of claim 1 wherein the determination of step (d) is done radiometrically.
Claim 12
Independent Claim
132 words
A method of determining the concentration of thyroid stimulating hormone in a human body fluid, the method comprising the steps of: (a) incubating the fluid with labeled antibodies to thyroid stimulating hormone under conditions sufficient to complex substantially all of the hormone in the fluid with the antibodies; (b) incubating the complexes of step (a) with a composite comprising antibodies to the hormone attached to a water insoluble carrier material, the incubation being under conditions sufficient to assure complexation of substantially all of the labeled complex of step (a) with the composite; (c) separating the incubation products of step (b) from the incubation medium; (d) determining the amount of label associated with the separated products; and (e) relating that determination to a standard to determine the concentration of the thyroid stimulating hormone.
Claim 13
Independent Claim
132 words
A method of determining the concentration of thyroid stimulating hormone in a human body fluid, the method comprising the steps of: (a) incubating the fluid with labeled antibodies to thyroid stimulating hormone under conditions sufficient to complex substantially all of the hormone in the fluid with the antibodies; (b) incubating the complexes of step (a) with a composite comprising antibodies to the hormone attached to a water insoluble carrier material, the incubation being under conditions sufficient to assure complexation of substantially all of the labeled complex of step (a) with the composite; (c) separating the incubation products of step (b) from the incubation medium; (d) determining the amount of label associated with the separated products; and (e) relating that determination to a standard to determine the concentration of the thyroid stimulating hormone.
Claim 14
Dependent Claim
17 words
The method of claim 11 wherein the labeled antibodies of step (a) are labeled with an enzyme.
Claim 15
Dependent Claim
18 words
The method of claim 11 wherein the labeled antibodies of step (a) are labeled with a fluorogenic material.
Claim 16
Dependent Claim
17 words
The method of claim 11 wherein the labeled antibodies of step (a) are labeled with a radioisotope.
Claim 17
Dependent Claim
18 words
The method of claim 11 wherein the composite of step (b) comprises antibodies immobilized on an organic carrier.
Claim 18
Dependent Claim
18 words
The method of claim 11 wherein the composite of step (b) comprises antibodies immobilized on an inorganic carrier.
Claim 19
Dependent Claim
11 words
The method of claim 16 wherein the carrier comprises glass particles.
Claim 20
Independent Claim
136 words
A method of detecting the presence of hepatitis associated antigen in a human body fluid, the method comprising the steps of: (a) incubating the fluid with labeled antibodies to the antigen, the incubation being under conditions sufficient to complex any antigen in the fluid with the labeled antibodies; (b) incubating the complex of step (a) with a composite comprising antibodies to the antigen immobilized on a water insoluble carrier material, the incubation being under conditions sufficient to complex at least a portion of the complex of step (a) with the composite; (c) separating the incubation products of step (b) from the incubation medium; (d) determining the amount of label associated with the separated incubation products; and (e) relating the determination of step (d) to a standard to determine if the antigen is present in the sample.
Claim 21
Independent Claim
136 words
A method of detecting the presence of hepatitis associated antigen in a human body fluid, the method comprising the steps of: (a) incubating the fluid with labeled antibodies to the antigen, the incubation being under conditions sufficient to complex any antigen in the fluid with the labeled antibodies; (b) incubating the complex of step (a) with a composite comprising antibodies to the antigen immobilized on a water insoluble carrier material, the incubation being under conditions sufficient to complex at least a portion of the complex of step (a) with the composite; (c) separating the incubation products of step (b) from the incubation medium; (d) determining the amount of label associated with the separated incubation products; and (e) relating the determination of step (d) to a standard to determine if the antigen is present in the sample.
Claim 22
Dependent Claim
17 words
The method of claim 18 wherein the labeled antibodies of step (a) are labeled with an enzyme.
Claim 23
Dependent Claim
18 words
The method of claim 18 wherein the labeled antibodies of step (a) are labeled with a fluorogenic material.
Claim 24
Dependent Claim
17 words
The method of claim 18 wherein the labeled antibodies of step (a) are labeled with a radioisotope.
Claim 25
Dependent Claim
22 words
The method of claim 18 wherein the composite of step (b) comprises antibodies to hepatitis associated antigen immobilized on an organic carrier.
Claim 26
Dependent Claim
22 words
The method of claim 18 wherein the composite of step (b) comprises antibodies to hepatitis associated antigens immobilized on an inorganic carrier.
Claim 27
Dependent Claim
12 words
The method of claim 23 wherein the inorganic carrier comprises glass particles.
Claim 28
Independent Claim
121 words
A method of determining the concentration of carcinoembryonic antigen in a body fluid, the method comprising the steps of: (a) incubating the fluid with labeled antibodies to carcinoembryonic antigen under conditions sufficient to complex the antigen with the antibodies; (b) incubating the complexed products of step (a) with a composite comprising antibodies to carcinoembryonic antigen immobilized on a water insoluble carrier material, the incubation being under conditions sufficient to assure complexation of the complex of step (a) with the composite; (c) separating the incubation products of step (b) from the incubation medium; (d) determining the amount of label associated with the separated products; and (e) relating the determination of step (d) to a standard to determine the concentration of the antigen.
Claim 29
Independent Claim
121 words
A method of determining the concentration of carcinoembryonic antigen in a body fluid, the method comprising the steps of: (a) incubating the fluid with labeled antibodies to carcinoembryonic antigen under conditions sufficient to complex the antigen with the antibodies; (b) incubating the complexed products of step (a) with a composite comprising antibodies to carcinoembryonic antigen immobilized on a water insoluble carrier material, the incubation being under conditions sufficient to assure complexation of the complex of step (a) with the composite; (c) separating the incubation products of step (b) from the incubation medium; (d) determining the amount of label associated with the separated products; and (e) relating the determination of step (d) to a standard to determine the concentration of the antigen.
Claim 30
Dependent Claim
17 words
The method of claim 25 wherein the labeled antibodies of step (a) are labeled with an enzyme.
Claim 31
Dependent Claim
18 words
The method of claim 25 wherein the labeled antibodies of step (a) are labeled with a fluorogenic material.
Claim 32
Dependent Claim
17 words
The method of claim 25 wherein the labeled antibodies of step (a) are labeled with a radioisotope.
Claim 33
Dependent Claim
21 words
The method of claim 25 wherein the composite of step (b) comprises antibodies to the antigen immobilized on an organic carrier.
Claim 34
Dependent Claim
21 words
The method of claim 25 wherein the composite of step (b) comprises antibodies to the antigen immobilized on an inorganic carrier.
Claim 35
Dependent Claim
11 words
The method of claim 30 wherein the carrier comprises glass particles.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-07-03 | 1982-01-02 | 1982-07-04 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-07-03 | 1986-01-02 | 1986-07-04 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-07-03 | 1990-01-02 | 1990-07-04 | $7700.00 | Window Closed / Lapsed |