System, device and method for high-throughput multi-plexed detection

Patent metadata

Field	Value
Publication number	US-12105086-B2
Application number	US-202117204279-A
Country	US
Kind code	B2
Filing date	Mar 17, 2021
Priority date	Aug 24, 2012
Publication date	Oct 1, 2024
Grant date	Oct 1, 2024

Abstract

The present invention relates to a system, device, and method for the high throughput multiplexed detection of a wide number of compounds. The invention comprises of a microwell array coupled to a capture agent array to form a plurality of interfaces between a microwell and a set of immobilized capture agents. The set of capture agents comprises a plurality of distinguishable features, with each feature corresponding to the detection of a particular compound of interest. In certain embodiments, each microwell is configured to contain a single cell. The invention is therefore capable of performing a high throughput analysis of single cell profiles, including profiles of secreted compounds.

First claim

What is claimed:

1. A device for the multiplexed detection of a plurality of compounds from single cells comprising two independent and separable components:
a microwell array comprising a plurality of individual microwells in uniform arrangement, at least some of the plurality of individual microwells having a length of greater than 50 μm and configured to contain an isolated single cell; and
a capture agent array comprising a plurality of immobilized capture agents, each immobilized capture agent capable of specifically binding to one of the plurality of compounds, where the immobilized capture agents are arranged in uniform capture agent sets, wherein each capture agent set comprises a plurality of isolated features at spatially identifiable locations, each isolated feature comprising at least one immobilized capture agent;
wherein the microwell array and capture agent array are separably coupled to form a plurality of enclosed interfaces, each enclosed interface comprising a microwell and a capture agent set such that the contents of each microwell are accessible to all of the isolated features of at least one set, thereby accessible to all of the immobilized capture agents.

2. The device of claim 1 , wherein each of the plurality of isolated features has a distinguishable spatial localization.

3. The device of claim 1 , wherein at least some of the plurality of microwells is a high aspect ratio rectangular well, having dimensions of about 1-2 mm in length and about 5-50 μm in depth.

4. The device of claim 1 , wherein the plurality of compounds comprise at least one protein secreted from a single cell contained within a microwell.

5. The device of claim 1 , wherein the plurality of capture agents comprise at least one molecule selected from the group consisting of an antibody, protein, peptide, peptide fragment, and nucleic acid.

6. The device of claim 1 , wherein each capture agent set comprises about 10-100 isolated features, each isolated feature comprising at least one immobilized capture agent that specifically binds to one compound.

7. The device of claim 1 , wherein the capture agent array comprises greater than 10 different capture agents, thereby allowing for the detection of greater than 10 different compounds.

8. A system for the multiplexed detection of a plurality of compounds from single cells comprising:
a device comprising two independent and separable components comprising:
a microwell array comprising a plurality of individual microwells in uniform arrangement, at least some of the plurality of individual microwells configured to contain a single cell; and
a capture agent array comprising a plurality of immobilized capture agents, each immobilized capture agent capable of specifically binding to one of the plurality of compounds, where the immobilized capture agents are arranged in uniform capture agent sets, wherein each capture agent set comprises a plurality of isolated features at spatially identifiable locations, each isolated feature comprising at least one immobilized capture agent;
wherein the microwell array and capture agent array are separably coupled to form a plurality of enclosed interfaces, each enclosed interface comprising a microwell and a capture agent set such that the contents of each microwell are accessible to all of the isolated features of at least one set, thereby accessible to all of the immobilized capture agents; and

a plurality of secondary capture agents, wherein each secondary capture agents comprises a detectable label and is configured to bind to an immobilized capture agent-compound complex formed at an isolated feature by the binding of a compound of the plurality of compounds to an immobilized capture agent of the plurality of immobilized capture agents.

9. The device of claim 1 , wherein at least one isolated feature contains at least two immobilized capture agents, and wherein each immobilized capture agent specifically recognizes a different compound of interest.

Assignees

• Univ Yale

Inventors

• Fan Rong
• Lu Yao
• Chen Jonathan

Classifications

• G01N2570/00

  Omics, e.g. proteomics, glycomics or lipidomics; Methods of analysis focusing on the entire complement of classes of biological molecules or subsets thereof, i.e. focusing on proteomes, glycomes or lipidomes · CPC title

• G01N33/54386

  Analytical elements · CPC title

• G01N33/54306

  Solid-phase reaction mechanisms · CPC title

• G01N2800/7028

  Cancer · CPC title

• G01N33/54366 Primary

  Apparatus specially adapted for solid-phase testing · CPC title