IP Library Granted Patent US 9,175,072
Granted Patent B2
US 9,175,072 · App. 13/374,351 · Granted Nov 3, 2015

Compositions and methods including recombinant B lymphocyte cell line including an exogenously incorporated nucleic acid expressing an exogenous membrane immunoglobulin reactive to a first antigen and including an endogenous gene expressing an endogenous secreted immunoglobulin reactive to a second antigen

Inventors: Roderick A. Hyde (Redmond, WA); Wayne R. Kindsvogel (Seattle, WA)
Assignee: Elwha LLC
C07K16/18A61K39/00C07K16/00C07K16/109C07K16/1018C07K16/1271C07K16/3069C07K16/44C12N5/0635G01N33/6854A61K2039/505A61K2039/5156C07K2317/14C07K2317/76C12N2510/00
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Quick Facts
Patent No.
US 9,175,072
App. No.
13/374,351
Granted
Nov 3, 2015
Kind
B2
Abstract

Compositions and methods are disclosed herein for producing one or more immunoglobulins in an isolated B lymphocyte cell line. An isolated cell line includes an isolated B lymphocyte cell line capable of expressing at least one exogenously incorporated membrane immunoglobulin reactive to a first antigen and at least one endogenous secreted immunoglobulin reactive to a second antigen.

Claims (56)

1. An isolated recombinant cell line comprising:

an isolated B lymphocyte cell line including at least one exogenously incorporated nucleic acid, the isolated B lymphocyte cell line capable of expressing the at least one exogenously incorporated nucleic acid encoding at least one exogenously membrane immunoglobulin reactive to a first antigen that is a B lymphocyte-activating antigen and at least one endogenous gene encoding at least one endogenous secreted immunoglobulin reactive to a second antigen from a pathogenic agent.

2. The isolated cell line of claim 1 comprising:

the isolated B lymphocyte cell line capable of expressing at least one endogenous membrane immunoglobulin reactive to the second antigen.

3. The isolated cell line of claim 1 , wherein the at least one exogenously incorporated membrane immunoglobulin comprises one or more exogenously incorporated membrane immunoglobulin polypeptides.

4. The isolated cell line of claim 1 , wherein the at least one exogenously incorporated membrane immunoglobulin comprises at least one exogenously incorporated nucleic acid encoding the at least one membrane immunoglobulin, wherein the cell line is encoding the at least one membrane immunoglobulin.

5. The isolated cell line of claim 4 , wherein the at least one exogenously incorporated membrane immunoglobulin comprises at least two exogenously incorporated nucleic acid encoding the at least one membrane immunoglobulin.

6. The isolated cell line of claim 5 , wherein the at least one exogenously incorporated membrane immunoglobulin comprises nucleic acids encoding two heavy chain (H) immunoglobulins and two light chain (L) immunoglobulins.

7. The isolated cell line of claim 5 , wherein the at least one exogenously incorporated membrane immunoglobulin comprises nucleic acids encoding one heavy chain (H) immunoglobulin and one light chain (L) immunoglobulin.

8. The isolated cell line of claim 4 , wherein the at least one exogenously incorporated membrane immunoglobulin comprises nucleic acids encoding one single chain Fv immunoglobulin.

9. The isolated cell line of claim 4 , wherein

the exogenously incorporated nucleic acid encoding the at least one membrane immunoglobulin is present in one or more chromosomal loci in the isolated B lymphocyte cell line.

10. The isolated cell line of claim 1 , wherein

the isolated B lymphocyte cell line is capable of disrupting expression of the endogenous membrane immunoglobulin reactive to the second antigen.

11. The isolated cell line of claim 5 , wherein

the at least two exogenously incorporated nucleic acids encoding the at least one of the membrane immunoglobulin are present in Ig H chain and Ig L chain chromosomal loci in the isolated B lymphocyte cell line.

12. The isolated cell line of claim 4 , wherein

the at least one exogenously incorporated nucleic acid encoding the at least one membrane immunoglobulin is present in one or more non-Ig L or non-Ig H chromosomal loci in the isolated B lymphocyte cell line.

13. The isolated cell line of claim 4 , wherein

the at least one exogenously incorporated nucleic acid encoding the at least one membrane immunoglobulin is present in an extrachromosomal replicating genetic element in the isolated B lymphocyte cell line.

14. The isolated cell line of claim 4 , wherein

the at least one exogenously incorporated nucleic acid encoding the at least one membrane immunoglobulin is derived from a B lymphocyte cell line.

15. The isolated cell line of claim 1 , wherein the at least one exogenously incorporated membrane immunoglobulin activated by the first antigen is capable of controlling expression of the at least one endogenous secreted immunoglobulin reactive to the second antigen.

16. The isolated cell line of claim 1 , wherein

the isolated B lymphocyte cell line comprises at least one of naïve B lymphocyte, immature B lymphocyte, transitional B lymphocyte, mature B lymphocyte, follicular B lymphocyte, memory B lymphocyte, plasmablast, or plasma cell.

17. The isolated cell line of claim 15 , wherein the isolated B lymphocyte cell line comprises a polyclonal population of B lymphocytes.

18. The isolated cell line of claim 15 , wherein the isolated B lymphocyte cell line comprises a monoclonal population of B lymphocytes.

19. The isolated cell line of claim 1 , wherein

the membrane immunoglobulin comprises at least one of a membrane anchor, an cytoplasmic domain, and an extracellular ligand-binding domain.

20. An isolated recombinant cell line comprising:

an isolated B lymphocyte cell line including at least one exogenously incorporated nucleic acid, the isolated B lymphocyte cell line capable of expressing the at least one exogenously incorporated nucleic acid encoding at least one exogenous membrane immunoglobulin reactive to a first antigen that is a B lymphocyte-activating antigen and the at least one exogenously incorporated nucleic acid encoding at least one exogenous secreted immunoglobulin reactive to a second antigen from a pathogenic agent.

21. The isolated cell line of claim 20 comprising:

the isolated B lymphocyte cell line capable of expressing at least one exogenously incorporated nucleic acid encoding membrane immunoglobulin reactive to the second antigen.

22. The isolated cell line of claim 20 comprising:

the isolated B lymphocyte cell line capable of expressing at least one exogenously incorporated nucleic acid encoding a secreted immunoglobulin reactive to a third antigen.

23. The isolated cell line of claim 22 , wherein the second antigen and the third antigen are different epitopes of a single antigenic polypeptide.

24. The isolated cell line of claim 20 , wherein the at least one exogenously incorporated membrane immunoglobulin comprises at least one exogenously incorporated membrane immunoglobulin polypeptide.

25. The isolated cell line of claim 20 , wherein the at least one exogenously incorporated membrane immunoglobulin comprises at least one exogenously incorporated nucleic acid encoding at least one membrane immunoglobulin polypeptide, wherein the cell line is capable of expressing the at least one membrane immunoglobulin polypeptide.

26. The isolated cell line of claim 25 , wherein the at least one exogenously incorporated nucleic acid encoding the at least one membrane immunoglobulin is present in one or more chromosomal loci in the isolated B lymphocyte cell line.

27. The isolated cell line of claim 26 , wherein the at least two exogenously incorporated nucleic acids encoding the at least one membrane immunoglobulins are present in Ig H chain and Ig L chain chromosomal loci in the isolated B lymphocyte cell line.

28. The isolated cell line of claim 26 , wherein

the at least one exogenously incorporated nucleic acids encoding the at least one membrane immunoglobulin is present in one or more non-Ig L or non-Ig H chromosomal loci in the isolated B lymphocyte cell line.

29. The isolated cell line of claim 28 , wherein

the at least one exogenously incorporated nucleic acids encoding the at least one membrane immunoglobulin is present in an extrachromosomal replicating genetic element in the isolated B lymphocyte cell line.

30. The isolated cell line of claim 20 , wherein

the nucleic acid encoding the at least one membrane immunoglobulin is derived from a B lymphocyte cell line.

31. The isolated cell line of claim 20 , wherein the at least one exogenously incorporated membrane immunoglobulin activated by the first antigen is capable of controlling expression of the at least one exogenously incorporated secreted immunoglobulin reactive to the second antigen.

32. The isolated cell line of claim 20 , wherein

the isolated B lymphocyte cell line comprises at least one of naïve B lymphocyte, immature B lymphocyte, transitional B lymphocyte, mature B lymphocyte, follicular B lymphocyte, memory B lymphocyte, plasmablast, or plasma cell.

33. The isolated cell line of claim 31 , wherein the isolated B lymphocyte cell line comprises a polyclonal population of B lymphocytes.

34. The isolated cell line of claim 31 , wherein the isolated B lymphocyte cell line comprises a monoclonal population of B lymphocytes.

35. The isolated cell line of claim 20 , wherein

the membrane immunoglobulin comprises at least one of a membrane anchor, a cytoplasmic domain, and an extracellular ligand-binding domain.

36. The isolated recombinant cell line of claim 1 , wherein the B lymphocyte-activating antigen is a model antigen, dinitrophenol, or keyhole limpet hemocyanin.

37. The isolated recombinant cell line of claim 1 , wherein the B lymphocyte-activating antigen is influenza antigen or S. aureus antigen.

38. The isolated recombinant cell line of claim 1 , wherein the disease-causing agent is an infectious disease-causing agent or a cancer-causing agent.

Assignments (4)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded May 6, 2021
From: KOTA BIOTHERAPEUTICS, LLC
To: THE INVENTION SCIENCE FUND II, LLC
Reel/Frame 056159/0064 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 13, 2019
From: THE INVENTION SCIENCE FUND II, LLC
To: KOTA BIOTHERAPEUTICS, LLC
Reel/Frame 050995/0470 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Aug 29, 2019
From: ELWHA LLC
To: THE INVENTION SCIENCE FUND II, LLC
Reel/Frame 050216/0254 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 13, 2012
From: HYDE, RODERICK A.; KINDSVOGEL, WAYNE R.
To: ELWHA LLC
Reel/Frame 027854/0193 →
Continuity (1)
Related Publication 20130164784A1 · Jun 27, 2013