Vectors used to create hybrid constant regions

• US 20070009957A1
• Filed: 03/04/2004
• Published: 01/11/2007
• Est. Priority Date: 03/04/2003
• Status: Abandoned Application

• Current Assignee: Alexion Pharmaceuticals Incorporated (Astrazeneca PLC)
• Original Assignee: Alexion Pharmaceuticals Incorporated (Astrazeneca PLC)
• Inventors: Frederickson, Shana, Maruyama, Toshiaki, Bowdish, Katherine
• Application Number:
  US10/547,275
• Publication Number:
  US 20070009957A1
• Time in Patent Office: Days
• US Class Current: 435/7.100
• CPC Class Codes: C07K 16/005 constructed by phage libraries
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/55 Fab or Fab'

Abstract

Expression vectors used for expression of mouse Fab libraries and expression of individual clones have portions of mouse constant regions replaced with human constant regions while maintaining desired cloning sites.

Claims

1. An expression vector comprising nucleic acid encoding at least an antibody light chain region and an antibody heavy chain region and restriction sites for direct cloning of murine antibody genes into the vector, at least one of the antibody light chain region or the antibody heavy chain region being a hybrid regidn which is at least in part murine and at least in part human.

2. A hybrid light chain in an expression vector as in claim 1 wherein the antibody light chain constant region comprises at least a portion of a human kappa light chain constant region.

3. An expression vector as in claim 1 wherein the heavy chain constant region comprises at least a portion of a human heavy chain constant region CH1 domain.

4. An expression vector as in claim 1 wherein the heavy chain constant region comprises at least a portion of a human heavy chain constant region hinge domain.

5. An expression vector as in claim 1 wherein both the antibody light chain region and the antibody heavy chain region are hybrid regions which are at least in part murine and at least in part human.

14. A library of antibodies produced using an expression vector in accordance with claim 1.

15. A method of screening an antibody library comprising providing a library of antibodies in accordance with claim 14 and selecting antibodies having desired properties from said library.

6. A method of Fab expression comprising:

transfecting a host cell with an expression vector that comprising nucleic acid encoding at least an antibody light chain region and an antibody heavy chain region, at least one of the antibody light chain region or the antibody heavy chain region being a hybrid region which is at least in part murine and at least in part human,

wherein expression of Fabs is increased compared to the same vector having a purely murine constant regions.

7. An antibody comprising a hybrid constant region that is at least in part murine and at least in part human.

8. An antibody as in claim 7 wherein the hybrid constant region is the heavy chain constant region.

9. An antibody as in claim 7 wherein the hybrid constant region is the light chain constant region.

10. An antibody as in claim 7 wherein both the light chain constant region and the heavy chain constant region are hybrid constant regions.

11. An antibody as in claim 7 wherein the hybrid constant region includes at least a portion of a human heavy chain constant region CH1 domain.

12. An antibody as in claim 7 wherein the hybrid constant region includes at least a portion of a human heavy chain constant region hinge domain.

13. An antibody as in claim 7 wherein the hybrid constant region includes at least a portion of a human kappa light chain constant region.

16. In a method of expressing Fabs wherein a host cells in transfected with an expression vector containing nucleic acid encoding at least a portion of a murine heavy chain variable region and at least a portion of a murine heavy chain constant region, the improvement comprising substituting nucleic acid encoding at least a portion of a human heavy chain constant region for a portion of the nucleic acid encoding at least a portion of a murine heavy chain constant region, wherein Fabs expressed contain a hybrid heavy chain constant region.

17. In a method of expressing Fabs wherein a host cells in transfected with an expression vector containing nucleic acid encoding at least a portion of a murine light chain variable region and at least a portion of a murine light chain constant region, the improvement comprising substituting nucleic acid encoding at least a portion of a human light chain constant region for a portion of the nucleic acid encoding at least a portion of a murine light chain constant region, wherein Fabs expressed contain a hybrid light chain constant region.

18. In a method of expressing Fabs wherein a host cells in transfected with an expression vector containing nucleic acid encoding at least a portion of a murine light chain variable region and at least a portion of a murine light chain constant region, the improvement comprising substituting nucleic acid encoding at least a portion of a human light chain constant region for a portion of the nucleic acid encoding at least a portion of a murine light chain constant region and substituting nucleic acid encoding at least a portion of a human heavy chain constant region for a portion of the nucleic acid encoding at least a portion of a murine heavy chain constant region, wherein Fabs expressed contain a hybrid light chain constant region and a hybrid heavy chain constraint region.