Methods and Compositions for Use in Analyte Detection Using Proximity Probes

Abstract

Methods and compositions for detecting an analyte in a sample are provided. In practicing the subject methods, a sample is combined with at least a pair of proximity probes that each include an analyte binding domain and a nucleic acid domain. The resultant mixture is then contacted with a pair of asymmetric nucleic acid connectors. Proximity dependent connector mediated interaction between the nucleic acid domains of the proximity probes is then detected to determine the presence of the analyte in the sample. Also provided are kits and systems for practicing the subject methods.

1 - 20 . (canceled) 
     
     
         21 . A kit for detecting and quantifying an analyte in a sample, said kit comprising:
 at least one set of proximity probes comprises at least a first and second proximity probe each comprising an analyte binding domain and a nucleic acid binding domain; and   first and second asymmetric connectors corresponding to said first and second proximity probes.   
     
     
         22 . A kit according to  claim 21 , further comprising a nucleic acid ligase. 
     
     
         23 . A kit according to  claim 21 , further comprising first and second supplementary nucleic acids. 
     
     
         24 . A kit according to  claim 21 , further comprising single strand binding proteins. 
     
     
         25 . A kit according to  claim 21 , further comprising PCR amplification reagents. 
     
     
         26 . A kit according to  claim 21 , wherein said analyte binding domain is an antibody or binding fragment thereof. 
     
     
         27 . A kit according to  claim 21 , wherein said kit comprises a single set of proximity probes. 
     
     
         28 . A kit according to  claim 21 , wherein said kit comprises two or more sets of proximity probes. 
     
     
         29 . A kit according to  claim 21 , wherein said first asymmetric connector comprises a first region of complementarity for said nucleic acid binding domain of said first proximity probe and a second region of complementarity for said nucleic acid binding domain of said second proximity probe, wherein said first region is longer than said second region. 
     
     
         30 . A kit according to  claim 21 , wherein said second asymmetric connector comprises a first region of complementarity for said nucleic acid binding domain of said second proximity probe and a second region of complementarity for said nucleic acid binding domain of said first proximity probe, wherein said first region is longer than said second region. 
     
     
         31 . The kit according to  claim 21 , wherein said kit further comprises a volume excluder. 
     
     
         32 . The kit according to  claim 31 , wherein said volume excluder is a macromolecule. 
     
     
         33 . The kit according to  claim 32 , wherein said macromolecule is a polymer. 
     
     
         34 . The kit according to  claim 33 , wherein said polymer is a PEG polymer.