Methods for conducting multiplexed assays

A method of conducting a multiplexed binding assay for a plurality of analytes comprising a first analyte of interest and a second analyte of interest comprising

• (a) combining, in one or more steps, the following components:
• (i) a sample comprising the plurality of analytes comprising the first analyte of interest and the second analyte of interest,
• (ii) a first targeting agent immobilized on a first binding domain, wherein the first targeting agent comprises an oligonucleotide,
• (iii) a first targeting agent complement connected to a first linking agent, wherein the first targeting agent complement comprises an oligonucleotide which is complementary to the oligonucleotide of the first targeting agent and forms a first oligonucleotide pair,
• (iv) a first binding reagent connected to a first supplemental linking agent, wherein the first binding reagent is a binding partner of the first analyte of interest,
• (v) a second targeting agent immobilized on a second binding domain, wherein the second targeting agent comprises an oligonucleotide,
• (vi) a second targeting agent complement connected to a second linking agent, wherein the second targeting agent complement comprises an oligonucleotide which is complementary to the oligonucleotide of the second targeting agent and forms a second oligonucleotide pair, and
• (vii) a second binding reagent connected to a second supplemental linking agent, wherein the second binding reagent is a binding partner of the second analyte of interest, wherein,
• the first linking agent binds to the first supplemental linking agent and the second linking agent binds to the second supplemental linking agent,
• (b) forming
• (i) a first binding complex on the first binding domain comprising the first targeting agent, the first targeting agent complement, the first binding reagent and the first analyte of interest, and
• (ii) a second binding complex on the second binding domain comprising the second targeting agent, the second targeting agent complement, the second binding reagent and the second analyte of interest, and
• (c) measuring the amount of the first analyte of interest and second analytes analyte of interest on the first binding domain and the second binding domain, respectively.