Assay methods

• US 9,618,510 B2
• Filed: 03/12/2014
• Issued: 04/11/2017
• Est. Priority Date: 03/13/2013
• Status: Active Grant

• Current Assignee: Meso Scale Technologies, LLC
• Original Assignee: Meso Scale Technologies, LLC
• Inventors: Aghvanyan, Anahit, Glezer, Eli N., Kenten, John, Sigal, George, Stengelin, Martin
• Primary Examiner(s): Gabel, Gail R
• Application Number:
  US14/206,284
• Publication Number:
  US 20140272939A1
• Time in Patent Office: 1,126 Days
• Field of Search: None
• CPC Class Codes: C12Q 1/6804 Nucleic acid analysis using immunogens immunoassay G01N33/53
• CPC Class Codes: C12Q 2537/125 Sandwich assay format
• CPC Class Codes: C12Q 2563/179 the label being a nucleic acid
• CPC Class Codes: G01N 2021/7786 Fluorescence
• CPC Class Codes: G01N 21/66 electrically excited, e.g. electroluminescence
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 21/77 by observing the effect on a chemical indicator
• CPC Class Codes: G01N 33/5306 Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/56988 HIV or HTLV
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

The present disclosure is directed to a kit for the detection of an analyte of interest in a sample comprising, in one or more vials, containers, or compartments, a. a surface comprising (i) a capture reagent for the analyte, said capture reagent bound to the surface, and (ii) an anchoring reagent bound to an anchoring oligonucleotide sequence, said anchoring reagent bound to the surface, b. a first detection reagent for the analyte that is linked to a first nucleic acid probe, wherein the first nucleic acid probe comprises an extended sequence that is complementary to the anchoring oligonucleotide sequence and c. a second detection reagent for the analyte that is linked to a second nucleic acid probe.

Claims

1. A kit for a sandwich binding assay detection of an analyte of interest in a sample comprising, in one or more vials, containers, or compartments:

a. a surface comprising (i) a capture reagent for the analyte, said capture reagent bound to the surface, and (ii) an anchoring reagent bound to an anchoring oligonucleotide sequence, said anchoring reagent bound to the surface;

b. a first detection reagent for the analyte that is linked to a first nucleic acid probe, wherein the first nucleic acid probe comprises an extended sequence that is complementary to the anchoring oligonucleotide sequence bound to the anchoring reagent; and

c. a second detection reagent for the analyte that is linked to a second nucleic acid probe;

wherein the kit is combined to form a proximity-based detection system.

2. The kit of claim 1 wherein the capture reagent comprises an antibody, antigen, ligand, receptor, oligonucleotide, hapten, epitope, mimitope, or aptamer.

3. The kit of claim 2 wherein the capture reagent comprises an antibody.

4. The kit of claim 1 wherein the first detection reagent comprises an antibody, antigen, ligand, receptor, oligonucleotide, hapten, epitope, mimitope, or aptamer.

5. The kit of claim 4 wherein the first detection reagent comprises an antibody.

6. The kit of claim 1 wherein the second detection reagent comprises an antibody, antigen, ligand, receptor, oligonucleotide, hapten, epitope, mimitope, or aptamer.

7. The kit of claim 1 wherein the second detection reagent comprises an antibody.

8. The kit of claim 1 wherein the surface comprises a particle.

9. The kit of claim 1 wherein the surface comprises a well of a multi-well plate.

10. The kit of claim 9 wherein the well comprises a plurality of distinct binding domains and the capture reagent and the anchoring reagent are bound to two distinct binding domains within the well.

11. The kit of claim 9 wherein the well comprises a plurality of distinct binding domains and the capture reagent and the anchoring reagent are bound to the same binding domain within the well.

12. The kit of claim 1 wherein the surface comprises a plurality of distinct binding domains and the capture reagent and the anchoring reagent are bound to two distinct binding domains on the surface.

13. The kit of claim 1 wherein the surface comprises a plurality of distinct binding domains and the capture reagent and the anchoring reagent are bound to the same binding domain on the surface.

14. The kit of claim 1 wherein the capture reagent and the anchoring reagent are bound to the surface within 10-100 nm of each other.

15. The kit of claim 1 wherein the surface comprises an electrode.