Nucleic acid linked immune-sandwich assay (NULISA)

An assay method for detecting one or multiple different analytes in a sample, comprising:

• (1) mixing one or more first binding moieties, wherein each of the one or more first binding moieties comprises a first binder and a first presenting group, one or more second binding moieties, wherein each of the one or more second binding moieties comprises a second binder and a second presenting group, and the sample in a solution, wherein for each pair of a first binding moiety and a second binding moiety:
• (i) the first binder and the second binder bind to an analyte of the one or multiple different analytes and form an immunocomplex,
• (ii) the immunocomplex is captured on a first solid surface in contact with the solution via binding between the first presenting group and a first receiving group coupled to the first solid surface, and
• (iii) the first binding moiety further comprises a first target label and the second binding moiety further comprises a second target label;
• (2) washing the first solid surface to remove unbound molecules;
• (2a) releasing the immunocomplex from the first solid surface;
• (2b) introducing a second solid surface and capturing the immunocomplex on the second solid surface via binding between the second presenting group and a second receiving group coupled to the second solid surface;
• (3) generating a reporter from the immunocomplex based on proximity between the first target label and the second target label;
• and
• (4) detecting the reporter, thereby detecting the analyte.