SANDWICH ASSAYS IN DROPLETS
The invention generally relates to performing sandwich assays in droplets. In certain embodiments, the invention provides methods for detecting a target analyte that involve forming a compartmentalized portion of fluid including a portion of a sample suspected of containing a target analyte and a sample identifier, a first binding agent having a target identifier, and a second binding agent specific to the target analyte under conditions that produce a complex of the first and second binding agents with the target analyte, separating the complexes, and detecting the complexes, thereby detecting the target analyte.
1 - 20 . (canceled)
21 . A method comprising:
incubating a plurality of target analytes with a plurality of binding agents, wherein binding agents of the plurality of binding agents comprise a binder and a target identifier comprising a barcode oligonucleotide sequence, thereby forming one or more complexes comprising a binding agent bound to a target analyte; and
detecting a barcode oligonucleotide sequence of a given complex of the one or more complexes to detect a target analyte in said given complex.
22 . The method of claim 21 , further comprising separating unbound binding agents from the one or more complexes.
23 . The method of claim 21 , wherein one or more barcode oligonucleotide sequences are amplified.
24 . The method of claim 23 , wherein the one or more barcode oligonucleotide sequences are amplified by rolling circle amplification.
25 . The method of claim 21 , wherein the binders of the plurality of binding agents are oligonucleotides.
26 . The method of claim 21 , wherein the detecting step uses hybridization from libraries of probes.
27 . The method of claim 21 , wherein one or more target analytes are labeled with an optically detectable label.
28 . The method of claim 27 , wherein the optically detectable label comprises a fluorescent label.
29 . The method of claim 21 , wherein the detecting step comprises sequencing the barcode oligonucleotide sequence of the given complex.