Unfolding proximity probes and methods for the use thereof

摘要

The present invention relates to a proximity-probe based detection assay for detecting an analyte in a sample and in particular to a method that comprises the use of at least one set of at least first and second proximity probes, which probes each comprise an analyte-binding domain and a nucleic acid domain and can simultaneously bind to the analyte directly or indirectly, wherein the nucleic acid domain of at least one of said proximity probes comprises a hairpin structure that can be unfolded by cleavage of the nucleic acid domain to generate at least one ligatable free end or region of complementarity to another nucleic acid molecule in said sample, wherein when the probes bind to said analyte unfolding said hairpin structure allows the nucleic acid domains of said at least first and second proximity probes to interact directly or indirectly.

主权项

1. A method of detecting an analyte comprising a proteinaceous molecule, a nucleic acid molecule, a cell, or a microorganism in a sample, the method comprising providing a set of at least first and second proximity probes, wherein the proximity probes each comprise an analyte-binding domain and a nucleic acid domain and simultaneously bind to the analyte directly or indirectly, wherein the nucleic acid domain of at least one of the proximity probes comprises a hairpin structure that can be unfolded by cleavage of the nucleic acid domain to generate at least one ligatable free end or region of complementarity to another nucleic acid molecule in the sample, contacting the proximity probes with the sample and binding the proximity probes to the analyte, cleaving the nucleic acid domain comprising the hairpin structure to unfold the hairpin structure, allowing the nucleic acid domains of the at least first and second proximity probes to interact directly or indirectly to form a ligation product or a nucleic acid extension product which is indicative of the presence of the analyte in the sample, and detecting the ligation product or the nucleic acid extension product, thereby to detect the analyte.