Bio-barcode based detection of target analytes

A method for detecting for the presence of one or more target analytes in a sample, each target analyte having at least two binding sites for specific binding interactions with specific binding complements, in a sample comprising the steps of:

• a) providing at least one type of capture substrates, the capture substrates having bound thereto at least one specific binding complement of the specific target analyte that binds to at least a first binding site of the specific target analyte;
• b) providing at least one type of microparticle detection probes, the microparticle detection probes comprising a microparticle having bound thereto (i) at least one specific binding complement to a specific target analyte and (ii) a plurality of DNA barcodes, the DNA barcodes are fluorescently labeled, wherein the specific binding complement bound to the microparticle detection probes binds to at least a second binding site of the target analyte;
• c) contacting the capture substrates with a sample believed to contain target analytes under conditions effective to allow for binding of the specific target analyte to the specific binding complement bound to the capture substrate so as to immobilize the target analytes onto the capture substrates;
• d) optionally washing the capture substrate to remove unbound materials;
• and
• e) contacting the immobilized target analytes with at least one type of microparticle detection probes under conditions effective to allow for binding between the target analytes and the specific binding complement bound to the microparticle detection probes to the analyte and to form a complex in the presence of the target analyte on the capture substrate;
• f) optionally isolating and washing the capture substrate to remove unbound microparticle detection probes;
• and
• g) detecting for the presence of a fluorescent signal from the DNA barcodes wherein the presence of a fluorescent signal is indicative of the presence of a specific target analyte in the sample.

A method for detecting for the presence of one or more target analytes in a sample, each target analyte having at least two binding sites for specific binding interactions with specific binding complements, in a sample comprising the steps of:

• a) providing at least one type of capture substrates, the capture substrates having bound thereto at least one specific binding complement of the specific target analyte that binds to at least a first binding site of the specific target analyte;
• b) providing at least one type of microparticle detection probes, the microparticle detection probes comprising a microparticle having bound thereto (i) at least one specific binding complement to a specific target analyte and (ii) a plurality of DNA barcodes, the DNA barcodes are fluorescently labeled, wherein the specific binding complement bound to the microparticle detection probes binds to at least a second binding site of the target analyte;
• c) contacting the capture substrates with a sample believed to contain target analytes under conditions effective to allow for binding of the specific target analyte to the specific binding complement bound to the capture substrate so as to immobilize the target analytes onto the capture substrates;
• d) optionally washing the capture substrate to remove unbound materials;
• and
• e) contacting the immobilized target analytes with at least one type of microparticle detection probes under conditions effective to allow for binding between the target analytes and the specific binding complement bound to the microparticle detection probes to the analyte and to form a complex in the presence of the target analyte on the capture substrate;
• f) optionally isolating and washing the capture substrate to remove unbound microparticle detection probes;
• g) subjecting the isolated washed capture substrate to conditions effective to release the DNA barcodes;
• and
• h) detecting for the presence of the fluorescent signal of the DNA barcodes wherein the presence of a fluorescent signal of the DNA barcode is indicative of the presence of a specific target analyte in the sample.

A method for detecting for the presence of one or more target analytes in a sample, each target analyte having at least two binding sites for specific binding interactions with specific binding complements, in a sample comprising the steps of:

• a) providing at least one type of capture substrates, the capture substrates having bound thereto at least one specific binding complement of the specific target analyte that binds to at least a first binding site of the specific target analyte;
• b) providing at least one type of microparticle detection probes, the microparticle detection probes comprising a microparticle having bound thereto (i) at least one specific binding complement to a specific target analyte and (ii) a plurality of DNA barcodes, the DNA barcodes comprising a reporter label, wherein the specific binding complement bound to the microparticle detection probes binds to at least a second binding site of the target analyte;
• c) contacting the capture substrates with a sample believed to contain target analytes under conditions effective to allow for binding of the specific target analyte to the specific binding complement bound to the capture substrate so as to immobilize the target analytes onto the capture substrates;
• d) optionally washing the capture substrate to remove unbound materials;
• and
• e) contacting the immobilized target analytes with at least one type of microparticle detection probes under conditions effective to allow for binding between the target analytes and the specific binding complement bound to the microparticle detection probes to the analyte and to form a complex in the presence of the target analyte on the capture substrate;
• f) optionally isolating and washing the capture substrate to remove unbound microparticle detection probes;
• g) subjecting the isolated washed capture substrate to conditions effective to release the DNA barcodes;
• and
• h) detecting for the presence of the reporter label of the DNA barcodes wherein the presence of the reporter label of the DNA barcode is indicative of the presence of a specific target analyte in the sample.

A method for detecting for the presence of one or more target analytes in a sample, each target analyte having at least two binding sites for specific binding interactions with specific binding complements, in a sample comprising the steps of:

• a) providing at least one type of capture substrates, the capture substrates having bound thereto at least one specific binding complement of the specific target analyte that binds to at least a first binding site of the specific target analyte;
• b) providing at least one type of particle detection probes, the particle detection probes comprising a particle having bound thereto (i) at least one specific binding complement to a specific target analyte and (ii) a plurality of DNA barcodes, the DNA barcodes are optionally labeled with a reporter label, wherein the specific binding complement bound to the particle detection probes binds to at least a second binding site of the target analyte;
• c) contacting the capture substrates with a sample believed to contain target analytes under conditions effective to allow for binding of the specific target analyte to the specific binding complement bound to the capture substrate so as to immobilize the target analytes onto the capture substrates;
• d) optionally washing the capture substrate to remove unbound materials;
• and
• e) contacting the immobilized target analytes with at least one type of particle detection probes under conditions effective to allow for binding between the target analytes and the specific binding complement bound to the particle detection probes to the analyte and to form a complex in the presence of the target analyte on the capture substrate;
• f) optionally isolating and washing the capture substrate to remove unbound particle detection probes;
• g) releasing the DNA barcodes from the particle detection probes by a chemical releasing agent;
• and
• h) detecting for the presence of the DNA barcode wherein the presence of the DNA barcode is indicative of the presence of a specific target analyte in the sample.

A method for detecting for the presence of one or more target analytes in a sample, each target analyte having at least two binding sites for specific binding interactions with specific binding complements, in a sample comprising the steps of:

• a) providing at least one type of capture substrates that are separable from the sample, the capture substrates having bound thereto at least one specific binding complement of the specific target analyte that binds to at least a first binding site of the specific target analyte;
• b) contacting the capture substrates with a sample believed to contain target analytes under conditions effective to allow for binding of the specific target analyte to the specific binding complement bound to the capture substrate so as to immobilize the target analytes onto the capture substrates;
• c) optionally separating the capture substrate and any target analytes bound thereto from the sample;
• d) providing at least one type of particle detection probes, the particle detection probes comprising a particle having bound thereto (i) at least one specific binding complement to a specific target analyte and (ii) a plurality of oligonucleotides that are optionally labeled with a reporter label, wherein the specific binding complement bound to the particle detection probes binds to at least a second binding site of the target analyte;
• e) contacting the immobilized target analytes with at least one type of particle detection probes under conditions effective to allow for binding between the target analytes and the specific binding complement bound to the particle detection probes to the analyte and to form a complex in the presence of the target analyte on the capture substrate, wherein the complex comprises the target analyte and capture substrate and particle detection probe;
• f) optionally separating the complex from unbound particle detection probes;
• and
• g) detecting for the presence of the complex wherein the presence of the complex is indicative of the presence of a specific target analyte in the sample.