Reverse sandwich immunoassay
First Claim
1. 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.
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Accused Products
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.
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Citations
31 Claims
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1. A method of determining the presence or concentration of a polyvalent antigenic substance in a fluid sample which comprises the steps of:
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(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. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A method of determining the concentration of thyroid stimulating hormone in a human body fluid, the method comprising the steps of:
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(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. - View Dependent Claims (12, 13, 14, 15, 16, 17)
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18. A method of detecting the presence of hepatitis associated antigen in a human body fluid, the method comprising the steps of:
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(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. - View Dependent Claims (19, 20, 21, 22, 23, 24)
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25. A method of determining the concentration of carcinoembryonic antigen in a body fluid, the method comprising the steps of:
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(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. - View Dependent Claims (26, 27, 28, 29, 30, 31)
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Specification