Screening assays and methods

Patent metadata

Field	Value
Publication number	US-10137426-B2
Application number	US-201615267977-A
Country	US
Kind code	B2
Filing date	Sep 16, 2016
Priority date	Sep 16, 2005
Publication date	Nov 27, 2018
Grant date	Nov 27, 2018

Abstract

Screening assays and methods of performing such assays are provided. In certain examples, the assays and methods may be designed to determine whether or not two or more species can associate with each other. In some examples, the assays and methods may be used to determine if a known antigen binds to an unknown monoclonal antibody.

First claim

What is claimed:

1. A method of producing a microarray of cell-derived products comprising:
(a) reversibly sealing a moldable slab to a surface of a substrate, said moldable slab comprising an array of microwells, each microwell having a side or diameter of less than 100 microns and containing not more than a single or a few cell(s), at least one of said microwells containing the single or few cell(s), wherein the single or few cell(s) produce a cell-derived product in a volume of 10 nanoliters or less of fluid in each said microwell containing the single or a few cell(s); wherein the surface of the substrate facing each said microwell is modified such that it acts to retain the cell-derived product on the surface of the substrate; and
(b) allowing said single or few cell(s) to produce said cell-derived product in said volume such that the cell-derived product produced by the single or few cell(s) in said volume is retained in a detectable amount on the surface of the substrate sealed to the moldable slab.

2. The method of claim 1 , wherein each microwell has a diameter of 50 to 100 microns.

3. The method of claim 1 , wherein the single or few cell(s) produce the cell-derived product in a volume of 100 picoliters to 1 nanoliter.

4. The method of claim 1 , wherein the substrate is a glass substrate.

5. The method of claim 1 , wherein the moldable slab is made of poly(dimethylsiloxane).

6. The method of claim 1 , wherein the cell-derived product is a cytokine, chemokine, inflammatory mediator, or growth factor.

7. The method of claim 1 , wherein the cell-derived product is a pathogen.

8. The method of claim 7 , wherein the pathogen is a virus.

9. The method of claim 1 , wherein the surface of the substrate is coated with an antibody that is capable of binding the cell-derived product.

10. The method of claim 1 , wherein the steps of reversibly sealing and allowing are performed in less than 24 hours.

11. The method of claim 1 , wherein the step of reversibly sealing forms a substantially fluid tight seal between the moldable slab and the substrate.

12. The method of claim 1 , wherein each microwell has a diameter of 10 to 100 microns; the single or few cell(s) produce the cell-derived product in a volume of 1 nanoliter or less; and the detectable amount is an amount suitable to detect association of a labeled species with said cell-derived product.

13. The method of claim 12 , wherein the step of reversibly sealing forms a substantially fluid tight seal between the moldable slab and the substrate.

14. The method of claim 13 , wherein the single or few cell(s) produce the cell-derived product in a volume of 100 picoliters to 1 nanoliter.

15. The method of claim 1 , wherein each microwell has a diameter of 10 to 100 microns; the single or few cell(s) produce the cell-derived product in a volume of 100 picoliters to 1 nanoliter; the detectable amount is an amount suitable to detect association of a fluorescently labeled species with said cell-derived product; the substrate is a glass substrate; the moldable slab is made of poly(dimethylsiloxane); and the step of reversibly sealing forms a substantially fluid tight seal between the moldable slab and the substrate.

16. The method of claim 1 , wherein the substrate is substantially planar.

17. The method of claim 12 , wherein the surface of the substrate facing said microwells is substantially planar.

18. The method of claim 1 , wherein the cell-derived product is secreted.

19. The method of claim 6 , wherein the cell-derived product is secreted.

20. The method of claim 1 , wherein the cell-derived product is liberated from the single or few cell(s) by lysis or permeabilization.

Assignees

• Harvard College

Inventors

• Love J Christopher
• Ploegh Hidde L
• Ronan Jehnna

Classifications

• B01L2200/12

  Specific details about manufacturing devices · CPC title

• B01L3/50853

  with covers or lids (closures for test tubes B01L3/50825) · CPC title

• G01N33/6854

  Immunoglobulins · CPC title

• B01J2219/0074

  Biological products · CPC title

• G01N33/505

  involving T-cells · CPC title