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

Patent metadata

Field	Value
Publication number	US-10274486-B2
Application number	US-201615290874-A
Country	US
Kind code	B2
Filing date	Oct 11, 2016
Priority date	Aug 24, 2012
Publication date	Apr 30, 2019
Grant date	Apr 30, 2019

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 multiplexed detection of molecules produced from a cell, the device comprising:
a plurality of enclosed interfaces, wherein each enclosed interface comprises
(a) a substrate containing at least ten spatially identifiable isolated features,
wherein of the at least ten of the isolated features, at least two isolated features contain at least two immobilized capture antibodies per isolated feature, and
wherein each immobilized capture antibody specifically recognizes a different protein of interest, and

(b) a chamber configured to contain a cell and the at least ten spatially identifiable isolated features of (a).

2. The device of claim 1 , wherein the chamber is a rectangle.

3. The device of claim 1 , wherein the chamber is a microchamber.

4. The device of claim 1 , wherein each isolated feature is separated from the next isolated feature by about 2-50 μm.

5. The device of claim 1 , wherein the isolated features are arrayed with a pitch size of 50 μm.

6. The device of claim 1 , wherein each isolated feature is 2-100 μm in width.

7. The device of claim 1 , wherein the enclosed interfaces are arranged in an array format including parallel lines or dots.

8. The device of claim 1 , wherein the isolated features are arranged in parallel within the enclosed interfaces.

9. The device of claim 3 , wherein the device comprises 200 microchambers per cm 2 to 20,000 microchambers per cm 2 .

10. The device of claim 3 , wherein the device comprises 100 microchambers to 100,000 microchambers.

11. The device of claim 3 , wherein each microchamber has a length of 10 μm to 2000 μm, a width of 10 μm to 100 μm, and a depth of 10 μm to 100 μm.

12. The device of claim 1 , wherein each of the isolated features contains at least three immobilized capture antibodies.

13. The device of claim 1 , wherein the isolated features are fabricated by a micro-scale printing technique.

14. The device of claim 13 , wherein the micro-scale printing technique is microspotting or inkjet printing.

15. The device of claim 1 , wherein the device comprises at least one cell within a chamber of at least one enclosed interface.

16. The device of claim 1 , wherein the chamber comprises proteins secreted from the at least one cell.

17. The device of claim 16 , wherein the proteins bind to one or more capture antibodies of at least one isolated feature of the substrate.

Assignees

• Univ Yale

Inventors

• Fan Rong
• Lu Yao
• Chen Jonathan

Classifications

• G01N2800/7028

  Cancer · CPC title

• G01N33/54366 Primary

  Apparatus specially adapted for solid-phase testing · CPC title

• G01N33/54306

  Solid-phase reaction mechanisms · CPC title

• 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