IP Library Granted Patent US 10,294,454
Granted Patent B2
US 10,294,454 · App. 15/245,584 · Granted May 21, 2019

Methods and kits for cell activation

Inventors: Ernest William Kovacs (Cohoes, NY); Anup Sood (Clifton Park, NY); Reginald Donovan Smith (Schenectady, NY); Evelina Roxana Loghin (Rexford, NY); Padmaparna Chadhuri (Bangalore, IN); Vandana Keskar (Niskayuna, NY); Chrystal Mae Chadwick (Latham, NY); Martin James Brown (Ballston Spa, NY)
Assignee: General Electric Company
C12N5/0636C07K16/2809C07K16/2818C12N2501/51C12N2501/515
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Quick Facts
Patent No.
US 10,294,454
App. No.
15/245,584
Granted
May 21, 2019
Kind
B2
Abstract

Provided are methods and kits for activating T cells, the method comprising providing a population of T cells, adding a plurality of first agents, where the first agent comprises a T-cell activator and a first binder moiety, and adding a second agent comprising a plurality of capture oligomers, where at least a segment of at least one of the plurality of capture oligomers is capable of associating with the first binder moiety. The method further comprises incubating the population of T cells, whereby at least a portion of the population of T cells is activated.

Claims (34)

1. A method of activating T cells, the method comprising:

a) providing a population of T cells;

b) adding a plurality of first agents, wherein the first agent comprises a T-cell activator attached to a first binder moiety;

c) adding a second agent comprising a plurality of capture oligomers, wherein at least a segment of at least one of the plurality of capture oligomers is capable of associating with the first binder moiety; and

d) incubating the population of T cells after steps (b) and (c), whereby at least a portion of the population of T cells is activated.

2. The method of claim 1 , wherein the second agent is a nucleic acid polymer comprising a plurality of capture oligonucleotide sequences.

3. The method of claim 2 , wherein the second agent is a rolling circle amplification product comprising a plurality of capture oligonucleotide sequences.

4. The method of claim 2 , wherein the first binder moiety is a nucleic acid sequence, wherein at least a segment of at least one of the plurality of capture oligonucleotide sequences is complementary to the nucleic acid sequence.

5. The method of claim 2 , further comprising adding a nuclease.

6. The method of claim 1 , wherein the T-cell activator is an anti-CD3 antibody or a fragment thereof.

7. The method of claim 1 , further comprising adding a T-cell co-stimulator.

8. The method of claim 7 wherein the T-cell co-stimulator is an anti-CD28 antibody, an anti-CD2 antibody or fragments thereof.

9. The method of claim 7 , wherein the T-cell co-stimulator is attached to a second binder moiety, wherein at least a segment of at least one of the plurality of capture oligomers is capable of associating with the second binder moiety.

10. The method of claim 9 , wherein the second binder moiety is a nucleic acid sequence.

11. The method according to claim 7 where the T-cell activator is an anti-CD3 antibody attached to a nucleic acid sequence, the T-cell co-stimulator is an anti-CD28 antibody attached to a nucleic acid sequence, and the second agent is a rolling circle amplification product comprising of a plurality of complementary capture oligonucleotide sequences.

12. A method according to claim 1 , further comprising a step of adding a vector comprising a foreign gene into the population of T cells.

13. A method according to claim 12 , wherein the vector is added with the plurality of first agents.

14. A method according to claim 12 , wherein the vector is added after at least a portion of the population of T cells is activated.

15. A method according to claim 12 , wherein the vector is a lentivirus vector or a gamma-retrovirus vector.

16. The method of claim 1 ,

wherein the T-cell activator is an anti-CD3 antibody, the first binder moiety is a nucleic acid sequences, the second agent is a nucleic acid polymer and the plurality of capture oligomers is a plurality of capture oligonucleotide sequences, wherein at least a segment of at least one of the plurality of capture oligonucleotide sequences is complementary to the nucleic acid sequence attached to the anti-CD3 antibody.

17. The method of claim 16 , further comprising adding an anti-CD28 antibody.

18. The method of claim 17 , wherein the anti-CD28 antibody is attached to a nucleic acid sequence, wherein at least a segment of at least one of the plurality of capture oligonucleotide sequences is complementary to the nucleic acid sequence attached to the anti-CD28 antibody.

19. The method of claim 2 , wherein the number of capture oligonucleotide sequences in the nucleic acid polymer is greater than 3.

20. The method of claim 2 , wherein the plurality of capture oligonucleotide sequences have the same nucleotide sequence or have different nucleotide sequences.

21. A method of activating cells, the method comprising:

a) providing a population of cells selected from the group consisting of B cells, T cells and natural killer cells;

b) adding a plurality of first agents, wherein the first agent comprises a cell activator attached to a first binder moiety:

c) adding a second agent comprising a plurality of capture oligomers, wherein at least a segment of at least one of the plurality of capture oligomers is capable of associating with the first binder moiety; and

d) incubating the population of cells after steps (b) and (c), whereby at least a portion of the population of cells is activated.

22. The method of claim 21 , wherein the second agent is a nucleic acid polymer comprising a plurality of capture oligonucleotide sequences.

23. The method of claim 22 , wherein the second agent is a rolling circle amplification product comprising a plurality of capture oligonucleotide sequences.

24. The method of claim 22 , wherein the first binder moiety is a nucleic acid sequence, wherein at least a segment of at least one of the plurality of capture oligonucleotide sequences is complementary to the nucleic acid sequence.

25. The method of claim 24 , wherein the cell activator is attached to the nucleic acid sequence via a covalent attachment.

Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Oct 1, 2020
From: GENERAL ELECTRIC COMPANY
To: GLOBAL LIFE SCIENCES SOLUTIONS USA LLC
Reel/Frame 053966/0133 →
CORRECTIVE ASSIGNMENT TO CORRECT THE FIFTH ASSIGNOR CHADHURI, PADMAPARNA PREVIOUSLY RECORDED ON REEL 039525 FRAME 0388. ASSIGNOR(S) HEREBY CONFIRMS THE ASSIGNMENT. Recorded Mar 25, 2019
From: KOVACS, ERNEST WILLIAM; SOOD, ANUP; SMITH, REGINALD DONOVAN; LOGHIN, EVELINA ROXANA; CHAUDHURI, PADMAPARNA; KESKAR, VANDANA; CHADWICK, CHRYSTAL MAE; BROWN, MARTIN JAMES
To: GENERAL ELECTRIC COMPANY
Reel/Frame 050719/0263 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 24, 2016
From: KOVACS, ERNEST WILLIAM; SOOD, ANUP; SMITH, REGINALD DONOVAN; LOGHIN, EVELINA ROXANA; CHADHURI, PADMAPARNA; KESKAR, VANDANA; CHADWICK, CHRYSTAL MAE; BROWN, MARTIN JAMES
To: GENERAL ELECTRIC COMPANY
Reel/Frame 039525/0388 →
Continuity (1)
Related Publication 20180057791A1 · Mar 1, 2018