Sensitive, multiplexed diagnostic assays for protein analysis
First Claim
1. A method for detecting multiple different compounds in a sample, said method comprising:
- (a) providing both a sample containing different compounds and a mixture of binding reagents, said binding reagents comprising nucleic acid-protein fusions, each fusion comprising (i) a protein portion which is known to specifically bind to one of said compounds and (ii) a nucleic acid portion which includes a unique identification tag;
(b) contacting said sample with said binding reagents under conditions allowing said protein portions of said binding reagents and said compounds to form binding reagent-compound complexes;
(c) capturing said binding reagent-compound complexes;
(d) amplifying said unique identification tag included on said nucleic acid portions of said binding reagent-compound complexes; and
(e) detecting said unique identification tag of each of said binding reagent-compound complexes, thereby detecting the corresponding compounds in said sample.
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Accused Products
Abstract
Disclosed herein are methods for detecting multiple compounds in a sample, involving: (a) contacting the sample with a mixture of binding reagents, the binding reagents being nucleic acid-protein fusions, each having (i) a protein portion which is known to specifically bind to one of the compounds and (ii) a nucleic acid portion which encodes the protein portion and which includes a unique identification tag; (b) allowing the protein portions of the binding reagents and the compounds to form complexes; (c) capturing the binding reagent-compound complexes; (d) amplifying the nucleic acid portions of the complexed binding reagents; and (e) detecting the unique identification tag of each of the amplified nucleic acids, thereby detecting the corresponding compounds in the sample. Also disclosed herein are kits for carrying out such methods.
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Citations
44 Claims
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1. A method for detecting multiple different compounds in a sample, said method comprising:
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(a) providing both a sample containing different compounds and a mixture of binding reagents, said binding reagents comprising nucleic acid-protein fusions, each fusion comprising (i) a protein portion which is known to specifically bind to one of said compounds and (ii) a nucleic acid portion which includes a unique identification tag;
(b) contacting said sample with said binding reagents under conditions allowing said protein portions of said binding reagents and said compounds to form binding reagent-compound complexes;
(c) capturing said binding reagent-compound complexes;
(d) amplifying said unique identification tag included on said nucleic acid portions of said binding reagent-compound complexes; and
(e) detecting said unique identification tag of each of said binding reagent-compound complexes, thereby detecting the corresponding compounds in said sample. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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13. The method of clam 12, wherein said solid support is a microchip.
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23. A kit for compound detection, said kit comprising:
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(a) a nucleic acid-protein fusion, wherein the protein portion of said fusion specifically binds said compound and the nucleic acid portion of said fusion includes a unique identification tag;
(b) a PCR primer pair, wherein the first primer hybridizes to said nucleic acid portion of said fusion 5′
to said unique identification tag and the second primer hybridizes to said nucleic acid portion of said fusion 3′
to said unique identification tag and hybridization of said first and second primers to said nucleic acid portion permits amplification of said unique identification tag; and
(c) a solid support comprising a nucleic acid which can hybridize to said unique identification tag. - View Dependent Claims (24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 37, 38, 39, 40, 41, 42, 43, 44)
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36. The method of clam 35, wherein said solid support is a microchip.
Specification