IP Library Granted Patent US 9,463,431
Granted Patent B2
US 9,463,431 · App. 14/928,607 · Granted Oct 11, 2016

Screening assays and methods

Inventors: J. Christopher Love (Somerville, MA); Hidde L. Ploegh (Brookline, MA); Jehnna Ronan (Chester, NH)
Assignee: President and Fellows of Harvard College
B01J19/0046B01L3/50853B82Y30/00G01N21/6452G01N33/505G01N33/531G01N33/543G01N33/54366G01N33/577G01N33/6854B01J2219/0074B01J2219/00675B01J2219/00722B01J2219/00725B01J2219/00734Y10S436/809Y10T436/24
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Quick Facts
Patent No.
US 9,463,431
App. No.
14/928,607
Granted
Oct 11, 2016
Kind
B2
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.

Claims (32)

1. An apparatus comprising a moldable slab reversibly sealed to a substrate surface, said moldable slab comprising an array of microwells, wherein each microwell 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 microwells contains 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 few cell(s); wherein the surface of the substrate facing each said microwell containing said single or few cell(s) is bound to the cell-derived product from said single or few cell(s) in their respective microwell.

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

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

4. The apparatus of claim 1 , wherein the single or few cell(s) produce the cell-derived product in a volume of 10 picoliters to 10 nanoliters.

5. The apparatus of claim 1 , wherein the single or few cell(s) produce the cell-derived product in a volume of 1 nanoliter or less.

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

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

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

9. The apparatus of claim 1 , wherein the single or few cell(s) are T cells or B cells.

10. The apparatus of claim 1 , wherein the single or few cell(s) are liver cells or kidney cells.

11. The apparatus of claim 1 , wherein the cell-derived product is DNA, RNA, or a lipid.

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

13. The apparatus of claim 1 , wherein the cell-derived product is a monokine or lymphokine.

14. The apparatus of claim 1 , wherein the cell-derived product is a pathogen.

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

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

17. The apparatus of claim 1 , wherein each microwell has a diameter of 10 to 100 microns; and the single or few cell(s) produce the cell-derived product in a volume of one nanoliter or less.

18. The apparatus of claim 17 , wherein the moldable slab is reversibly sealed to the substrate so as to form a substantially fluid tight seal.

19. The apparatus of claim 18 , wherein the single or few cell(s) produce the cell-derived product in a volume of 100 picoliters to 1 nanoliter.

20. The apparatus 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 substrate is a glass substrate;

the moldable slab is made of poly(dimethylsiloxane); and

the moldable slab is reversibly sealed to the substrate so as to form a substantially fluid tight seal.

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

22. The apparatus of claim 1 , wherein the substrate is substantially planar.

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

24. The apparatus of claim 1 , wherein the cell-derived product is secreted.

25. The apparatus of claim 12 , wherein the cell-derived product is secreted.

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

27. The apparatus of claim 11 , wherein the cell-derived product is liberated from the single or few cell(s) by lysis or permeabilization.

28. The apparatus of claim 14 , wherein the pathogen is a virus.

Assignments (2)
MERGER Recorded Apr 9, 2026
From: BRUKER CELLULAR ANALYSIS, INC.
To: BRUKER SPATIAL BIOLOGY, INC.
Reel/Frame 074322/0723 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 3, 2015
From: LOVE, CHRISTOPHER; PLOEGH, HIDDE L.; RONAN, JEHNNA
To: PRESIDENT AND FELLOWS OF HARVARD COLLEGE
Reel/Frame 036945/0951 →
Continuity (4)
Continuation 12857508 · Aug 16, 2010
Division 11523124 · Sep 18, 2006
Provisional Application 60717976 · Sep 16, 2005
Related Publication 20160045885A1 · Feb 18, 2016