AUTOANTIBODY ENHANCED IMMUNOASSAYS AND KITS
First Claim
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1. An immunoassay for detecting an analyte of interest in a test sample, the immunoassay comprising the steps of:
- a) contacting a test sample suspected of containing an analyte of interest with a first antibody that binds to at least one epitope on the analyte of interest to form a first antibody-analyte complex, wherein the first antibody is immobilized on a solid phase, and further wherein at least one autoantibody in the test sample binds to at least one epitope on the analyte of interest to form an autoantibody-analyte complex, wherein said autoantibody binds to the solid phase;
b) contacting said mixture comprising a first antibody-analyte complex and an autoantibody-analyte complex with a second antibody to form a measurable assembly comprising a first antibody-analyte-second antibody complex and an autoantibody-analyte-second antibody complex;
wherein the second antibody binds to at least one epitope on the analyte of interest, and further wherein, an optical, electrical, or change-of-state signal of the assembly is measured.
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Abstract
The present disclosure provides immunoassays and kits for detection or quantification of an analyte of interest in a test sample that potentially contains endogenously produced autoantibodies reactive with the analyte.
26 Citations
35 Claims
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1. An immunoassay for detecting an analyte of interest in a test sample, the immunoassay comprising the steps of:
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a) contacting a test sample suspected of containing an analyte of interest with a first antibody that binds to at least one epitope on the analyte of interest to form a first antibody-analyte complex, wherein the first antibody is immobilized on a solid phase, and further wherein at least one autoantibody in the test sample binds to at least one epitope on the analyte of interest to form an autoantibody-analyte complex, wherein said autoantibody binds to the solid phase; b) contacting said mixture comprising a first antibody-analyte complex and an autoantibody-analyte complex with a second antibody to form a measurable assembly comprising a first antibody-analyte-second antibody complex and an autoantibody-analyte-second antibody complex;
wherein the second antibody binds to at least one epitope on the analyte of interest, and further wherein, an optical, electrical, or change-of-state signal of the assembly is measured. - View Dependent Claims (2, 3, 4, 5)
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6. An immunoassay for detecting an analyte of interest in a test sample, the immunoassay comprising the steps of:
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a) contacting a test sample suspected of containing an analyte of interest with a first antibody that binds to at least one epitope on the analyte of interest to form a first antibody-analyte complex, wherein the first antibody is immobilized on a solid phase, and further wherein at least one autoantibody in the test sample binds to at least one epitope on the analyte of interest to form an autoantibody-analyte complex, wherein said autoantibody binds to the solid phase; b) contacting said mixture comprising a first antibody-analyte complex and an autoantibody-analyte complex, with a second antibody that has been conjugated to a detectable label to form a first antibody-analyte-second antibody complex and an autoantibody-analyte-second antibody complex;
wherein the second antibody binds to at least one epitope on the analyte of interest and further wherein, the detectable label is at least one acridinium compound;c) generating or providing a source of hydrogen peroxide to the mixture of step (b); d) adding a basic solution to the mixture of step (c) to generate a light signal; and e) measuring the light signal generated by or emitted in step (d) and detecting the analyte of interest in the test sample. - View Dependent Claims (7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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20. A kit for detecting or quantifying an analyte of interest in a test sample, the kit comprising:
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a) a solid phase capable of binding autoantibodies present in the test sample; b) a first antibody that binds to at least one epitope on the analyte of interest, the first antibody bound to the solid phase; c) a second antibody that binds to at least one epitope on the analyte of interest; and d) instructions for detecting or quantifying the analyte of interest. - View Dependent Claims (21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33)
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34. An immunodetection composition comprising:
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a) a first detection complex comprising a first antibody reactive with an analyte of interest and bound to a solid phase, the analyte of interest, and a second antibody reactive with the analyte of interest, wherein the second antibody has a detectable label; and b) a second detection complex comprising an autoantibody reactive with the analyte of interest and bound to the solid phase, the analyte of interest, and the second antibody, wherein the first and second complexes generate a measurable optical, electrical, or change-of-state signal from the detectable label. - View Dependent Claims (35)
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Specification