Dioxetane-nanoparticle assemblies for energy transfer detection systems, methods of making the assemblies, and methods of using the assemblies in bioassays

An assay comprising:

• combining a chemiluminescent compound, a nanoparticle comprising an inorganic semiconductor material and a sample comprising an analyte in aqueous solution;
• wherein the analyte comprises an enzyme;
• and
• wherein the chemiluminescent compound can be activated by the enzyme to produce an excited state donor, wherein the excited state donor associates with the nanoparticle such that the excited state donor transfers energy via a non-radiative process to the inorganic semiconductor material of the nano-particle which thereby produces photoluminescent emissions.

An enzyme substrate comprising:

• a nanoparticle comprising an inorganic semiconductor material;
• and
• at least one molecule of a chemiluminescent compound associated with the nanoparticle;
• wherein the chemiluminescent compound can be activated by an enzyme to produce an excited state donor;
• and
• wherein the excited state donor transfers energy via a non-radiative process to the inorganic semiconductor material which thereby produces photoluminescent emissions.

An assay comprising:

• contacting a sample comprising a first analyte and a second analyte with a first substrate, the first substrate comprising a nanoparticle of an inorganic semiconductor material and at least one molecule of a first chemiluminescent compound associated with the nanoparticle, wherein the first chemiluminescent compound can be activated by a first enzyme to produce an excited state donor which transfers energy in a non-radiative process to the inorganic semiconductor material thereby producing a first photoluminescent emission and wherein the first analyte is capable of activating the first chemiluminescent compound;
• contacting the sample with a second substrate, the second substrate comprising a nanoparticle of an inorganic semiconductor material and at least one molecule of a second chemiluminescent compound associated with the nanoparticle, wherein the second chemiluminescent compound can be activated by a second enzyme to produce an excited state donor which transfers energy in a non-radiative process to the nanoparticle thereby producing a second photoluminescent emission spectrally distinct from the first photoluminescent emission and wherein the second analyte is capable of activating the second chemiluminescent compound;
• detecting the first and second photoluminescent emissions;
• wherein the first photoluminescent emission indicates the presence and/or amount of the first analyte in the sample, and wherein the second photoluminescent emission indicates the presence and/or amount of the second analyte in the sample.

An assay for detecting the presence of a first and a second analyte in a sample comprising:

• contacting the sample with a first substrate, the first substrate comprising a nanoparticle, at least one molecule of a first chemiluminescent compound associated with the nanoparticle, and at least one molecule of a first capture agent associated with the nanoparticle, wherein the first capture agent can associate with the first analyte, wherein the first chemiluminescent compound can be activated by a first enzyme to produce an excited state donor which transfers energy in a non-radiative process to the nanoparticle thereby producing a first photoluminescent emission;
• contacting the sample with a second substrate, the second substrate comprising a nanoparticle, at least one molecule of a second chemiluminescent compound associated with the nanoparticle, and at least one molecule of a second capture agent associated with the nanoparticle, wherein the second capture agent can associate with the second analyte, wherein the second chemiluminescent compound can be activated by a second enzyme to produce an excited state donor which transfers energy in a non-radiative process to the nanoparticle thereby producing a second photoluminescent emission spectrally distinct from the first photoluminescent emission;
• allowing the first analyte in the sample to associate with the first capture agent and allowing the second analyte in the sample to associate with the second capture agent;
• and
• detecting the first and second photoluminescent emissions;
• wherein the first photoluminescent emission indicates the presence and/or amount of the first analyte in the sample, and wherein the second photoluminescent emission indicates the presence and/or amount of the second analyte in the sample.