Screening assays and methods

Patent metadata

Field	Value
Publication number	US-11154833-B2
Application number	US-201816170323-A
Country	US
Kind code	B2
Filing date	Oct 25, 2018
Priority date	Sep 16, 2005
Publication date	Oct 26, 2021
Grant date	Oct 26, 2021

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. An apparatus comprising a moldable slab in contact with a substrate, said moldable slab defining a plurality of wells, wherein each well has a side or diameter of less than 100 microns and contains not more than a single or a few cell(s), wherein at least one of said wells contains the respective single or few cell(s), said respective single or few cell(s) having produced one or more cell-derived product(s) in a volume no greater than about 10 nanoliters in each said well containing said respective single or few cell(s), said one or more cell-derived product(s) being bound to the substrate at a location in fluid communication with said respective well containing said respective single or few cell(s).

2. The apparatus of claim 1 , wherein each well has a diameter of 10 to 100 microns.

3. The apparatus of claim 2 , wherein each well has a diameter of 50 to 100 microns.

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

5. The apparatus of claim 1 , wherein the substrate is a glass substrate.

6. The apparatus of claim 1 , wherein the moldable slab comprises poly(dimethylsiloxane).

7. The apparatus of claim 1 , wherein the respective single or few cell(s) comprise(s) one or more members selected from the group consisting of a T cells, a B cells, a liver cells, and a kidney cells.

8. The apparatus of claim 1 , wherein the cell-derived product comprise(s) one or more members selected from the group consisting of a DNA, RNA, cytokine, chemokine, inflammatory mediator, monokine, lymphokine, a growth factor, a lipid, and a pathogen.

9. The apparatus of claim 1 , wherein the moldable slab is reversibly sealed to the substrate so as to form a substantially fluid tight seal.

10. The apparatus of claim 1 , wherein each well has a diameter of 10 to 100 microns;
the respective single or few cell(s) produce the cell-derived product in a volume of 100 picoliters to 1 nanoliter;
the substrate is a glass substrate;
the moldable slab comprises poly(dimethylsiloxane); and
the moldable slab is reversibly sealed to the substrate so as to form a substantially fluid tight seal.

11. The apparatus of claim 1 , wherein the substrate comprises a plastic material.

12. The apparatus of claim 1 , wherein the cell-derived product(s) is/are secreted.

13. The apparatus of claim 1 , wherein the cell-derived product(s) is/are liberated from the respective single or few cell(s) by lysis or permeabilization.

14. The apparatus of claim 1 , wherein the substrate has a capture ligand immobilized thereon for retaining at least one of the cell-derived product(s), wherein the capture ligand is an antigen, a protein or an antibody.

15. The apparatus of claim 1 , wherein the substrate has a substantially planar surface.

16. A method comprising temporarily confining cells in a plurality of wells using the apparatus of claim 1 , and selectively recovering the single or few cell(s) from an identified well, said well identified from the one or more cell-derived product(s) produced by said single or few cell(s) while temporarily confined in said well, said one or more cell-derived product(s) being bound to the substrate at a location having been in fluid communication with said well.

Assignees

• Harvard College

Inventors

• Love J Christopher
• Ploegh Hidde L
• Ronan Jehnna

Classifications

• B01L2300/12

  Specific details about materials · CPC title

• B01L2200/12

  Specific details about manufacturing devices · CPC title

• B01L2200/025

  Align devices or objects to ensure defined positions relative to each other · CPC title

• B01L2300/0819

  Microarrays; Biochips · CPC title

• B01J2219/00725

  Peptides · CPC title