Method and device for immunoassay using nucleotide conjugates
First Claim
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1. A method of determining the presence of an analyte in a blood sample comprising:
- contacting the blood sample with a first antibody such that the first antibody becomes solubilized in the blood sample and binds to the analyte, wherein the first antibody is bounded to at least one signal enzyme via a synthetic nucleotide bridge comprising a first single stranded nucleotide and second single stranded nucleotide;
contacting the blood sample comprising the solubilized first antibody and the antigen with an immobilized second antibody such that the immobilized second antibody binds to the antigen to form an immunoassay;
washing the blood sample from the immunoassay; and
determining the presence of the analyte based on a signal generated by a reaction with the at least one signal enzyme,wherein a 3′
-end or 5′
-end of the a first single stranded nucleotide and the a second single stranded nucleotide is protected from endogenous exonuclease activity from said blood sample contacting said second antibody by incorporating a protective chemical group.
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Abstract
A composition of matter for use in an immunoassay devices and method comprising a signal antibody, e.g., FAB fragment, covalently linked to a first nucleotide; and one or more signal elements, e.g., signal enzymes such as ALP or fluorescent dyes, each covalently linked to a second nucleotide, wherein the first nucleotide has one or more repeated sequences, and the second nucleotide is bound to one of the one or more repeated sequences on said first nucleotide, and wherein the ratio of the signal antibody to the signal element is controlled by the number of repeated sequences.
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9 Claims
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1. A method of determining the presence of an analyte in a blood sample comprising:
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contacting the blood sample with a first antibody such that the first antibody becomes solubilized in the blood sample and binds to the analyte, wherein the first antibody is bounded to at least one signal enzyme via a synthetic nucleotide bridge comprising a first single stranded nucleotide and second single stranded nucleotide; contacting the blood sample comprising the solubilized first antibody and the antigen with an immobilized second antibody such that the immobilized second antibody binds to the antigen to form an immunoassay; washing the blood sample from the immunoassay; and determining the presence of the analyte based on a signal generated by a reaction with the at least one signal enzyme, wherein a 3′
-end or 5′
-end of the a first single stranded nucleotide and the a second single stranded nucleotide is protected from endogenous exonuclease activity from said blood sample contacting said second antibody by incorporating a protective chemical group. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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Specification