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.
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Citations
19 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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Specification