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

Patent metadata

Field	Value
Publication number	US-9506917-B2
Application number	US-201514629164-A
Country	US
Kind code	B2
Filing date	Feb 23, 2015
Priority date	Aug 24, 2012
Publication date	Nov 29, 2016
Grant date	Nov 29, 2016

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 compounds secreted from a cell, the device comprising:
a plurality of enclosed interfaces, wherein each enclosed interface comprises (a) a microchamber configured to contain a cell and (b) a substrate containing at least ten spatially identifiable isolated features, wherein at least ten of the isolated features contain at least two immobilized capture antibodies per isolated feature, and wherein each immobilized capture antibody recognizes a different protein of interest.

2. The device of claim 1 , wherein at least ten of the isolated features contain at least three immobilized capture antibodies per isolated feature.

3. The device of claim 1 , wherein the enclosed interfaces are arranged in parallel.

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

5. The device of claim 1 , wherein the device comprises 200 microchambers per cm 2 to 20,000 microchambers per cm 2 .

6. The device of claim 1 , wherein the device comprises 100 microchambers to 100,000 microchambers.

7. The device of claim 1 , wherein at least one of the isolated features on the substrate comprises 5 to 100 different capture antibodies.

8. The device of claim 1 , wherein each microchamber has a length of 100 μm to 2000 μm, a width of 10 μm to 100 μm, and a depth of 10 μm to 100 μm.

9. The device of claim 1 , wherein each isolated feature has a width of 5 μm to 50 μm.

10. The device of claim 1 , wherein the isolated features are separated from each other by 5 μm to 50 μm.

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

12. The device of claim 1 , wherein the isolated feature is a microchannel.

13. The device of claim 1 , wherein the isolated features are fabricated by a cross-flow patterning technique.

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

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

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

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

18. The device of claim 17 , wherein the proteins are bound to capture antibodies of isolated features of the substrate.

Assignees

• Univ Yale

Inventors

• Fan Rong
• Lu Yao
• Chen Jonathan

Classifications

• G01N33/54306 Primary

  Solid-phase reaction mechanisms · CPC title

• G01N33/54366 Primary

  Apparatus specially adapted for solid-phase testing · CPC title

• G01N2800/7028

  Cancer · 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