DNA-CONJUGATED ANTIBODIES FOR IMPROVED ANTIBODY AFFINITY AND REDUCED ANTIBODY CROSS REACTIVITY

摘要

Disclosed are specific binding molecules (e.g. antibodies) that are provided in a set of conjugates, where each conjugate comprises an antibody linked to an oligonucleotide (oligo), such as a DNA oligonucleotide. The antibodies in each set are to the same target antigen. One antibody is preferably immobilized so that it remains in the sample reaction after a wash step. One antibody is used for detection since it will remain in the sample after washing only if there is a specific antibody-target antigen. The oligos in a given set hybridize to each other when both antibodies bind to a target with immuno-specificity. However, if the binding is not immune specific, such as in the case of cross-reactivity, the oligos do not hybridize.

主权项

1. A method for detecting a target antigen in a sample, using a first antibody and a second antibody which can concurrently bind to the target antigen, comprising:
(a) providing a first antibody linked to a first polynucleic acid strand and a second antibody linked to a second polynucleic acid strand, wherein the first polynucleic acid strand and the second polynucleic acid strand comprise complementary portions that permit hybridization and formation of a polynucleotide complex between the first polynucleic acid strand and the second polynucleic acid strand when the first antibody is in proximity to the second antibody as a result of binding to the target antigen; (b) forming, in the sample, a complex wherein a target antigen may be bound to said first antibody and also bound to said second antibody; (c) incubating the complex of step (b) under conditions which allow polynucleic acid strands of step (a) to hybridize and form a polynucleotide complex between the first polynucleic acid strand and the second polynucleic acid strand; (d) subjecting the complex in step (c) to a wash step which removes any nonspecifically bound antibody from the target antigen but does not remove any specifically bound antibody, whereby polynucleotide hybridization occurs; and (e) detecting a presence in the sample of an antibody after the wash of step (d), wherein antibody not having a hybridizing strand is removed in step (d) and wherein presence of the antibody in step (e) having a hybridizing strand indicates specific binding to the target antigen.