Homogeneous analyte detection
The present invention provides novel binding pair compositions of defined and limited stability comprising nucleic acid detection markers useful for the homogeneous, sensitive detection of analytes. Also provided are methods for the sensitive homogeneous detection of analytes, particularly analytes of clinical relevance. Kits for preparing binding pairs of the invention and for performing the methods of the invention are also provided.
1. A binding pair comprising:
a first binding member comprising a first antibody to an analyte coupled to the 5′ end of a first single-stranded nucleic acid; and
a second binding member comprising a second antibody to the analyte coupled to the 3′ end of a second single-stranded nucleic acid, wherein the first antibody and second antibody form a complex with the analyte, and further wherein the first and second single-strand nucleic acids of the complex form a duplex of defined and limited stability between the terminal 3′ end of the first nucleic acid and an internal sequence of the second nucleic acid; and wherein the duplex of defined and limited stability can be dissociated while the antibodies remain in place in close proximity through binding with the analyte and then reassociated to form a duplex suitable for amplification.
2. The binding pair of claim 1 , wherein the first and second antibodies are monoclonal antibodies.
3. The binding pair of claim 2 , wherein the first antibody and the second antibody interact independently with epitopes on the same antigen.
4. The binding pair of claim 2 , wherein the first antibody and the second antibody comprise a sandwich pair.
5. The binding pair of any one of claims 1 , 2 , 3 or 4 , wherein the first nucleic acid comprises SEQ ID NO. 1 and the second nucleic acid comprises SEQ ID NO.:2 or SEQ ID NO.:3.
6. The binding pair of any one of claims 1 , 2 , 3 or 4 , wherein the first and second nucleic acids are each independently selected from the group consisting of DNA, RNA, and PNA.
7. The binding pair of any one of claim 1 , 2 , 3 or 4 , wherein at least one of the first and second nucleic acids is a chimeric DNA/RNA molecule.
8. The binding pair of claim 1 , wherein the nucleic acid hybrid is suitable for amplification by PCR, LCR, SDA, or TMA.
9. The binding pair of claim 1 , wherein the duplex of defined and limited stability can be dissociated at a temperature between 26° C. and 45° C. while the antibodies remain in place in close proximity through binding with the analyte.