Reverse sandwich immunoassay
AssigneeCorning Glass Works
InventorsRoger N. Piasio, James W. Ryan, James E. Woiszwillo
Life of this patent
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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.
Claims31
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.
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.
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.
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.
Dependencies are inferred from the claim text.