Nucleic acid encoding antibodies binding to the extracellular domain of the receptor tyrosine kinase ALK
View Patent ↗The present invention concerns an antibody specific for human ALK (Anaplastic Lymphoma Kinase), in particular a scFv, a nucleic acid sequence encoding it, its production and its use as a pharmaceutical or for diagnostic purposes. Said antibody is suitable for the local treatment of tumors, in particular glioblastoma.
1. An isolated nucleic acid encoding a monoclonal antibody or antigen binding fragment thereof, wherein said antibody or fragment specifically binds human ALK protein, said antibody comprising a heavy chain variable region (VH) domain and a light chain variable region (VL) domain, said heavy chain variable region domain comprising the 3 CDRs in SEQ ID NO: 4 and said light chain variable region domain comprising the 3 CDRs in SEQ ID NO:5.
2. An expression vector comprising the isolated nucleic acid of claim 1 .
3. A suitable host cell transformed with an expression vector of claim 2 , in particular an E. coli cell.
4. A method for the production of an antibody comprising culturing the host cell of claim 3 under conditions that allow the synthesis of said antibody and recovering said antibody.
5. The isolated nucleic acid of claim 1 , wherein the monoclonal antibody or fragment is selected from the group consisting of a single chain antibody (scFv), a Fab fragment, an IgG antibody and an IgM antibody.
6. The isolated nucleic acid of claim 1 , said antibody or fragment having a binding affinity for the ALK epitope characterized by a K d of 30 nM or less.
7. The isolated nucleic acid of claim 1 , said antibody or fragment having a binding affinity for the ALK epitope characterized by a K d of less than 3 nM.
8. The isolated nucleic acid of claim 1 , wherein the VH and VL domains comprise frameworks that are stable and soluble so as to be functional in an intracellular reducing environment.
9. The isolated nucleic acid of claim 1 , derived from framework 4.4 (SEQ ID NO: 20).
10. The isolated nucleic acid of claim 1 , wherein the VH domain is a H3 VH domain and the VL domain is a lambda1 VL domain.
11. An isolated nucleic acid encoding a monoclonal antibody or antigen binding fragment thereof, wherein said antibody or fragment specifically binds human ALK protein, said antibody comprising a heavy chain variable region domain comprising the sequence of SEQ ID NO:4 and a light chain variable region domain comprising the sequence of SEQ ID NO:5.
12. The isolated nucleic acid of claim 11 , wherein the monoclonal antibody or fragment is selected from the group consisting of a single chain antibody (scFv), a Fab fragment, an IgG antibody and an IgM antibody.
13. The isolated nucleic acid of claim 11 , said antibody or fragment having a binding affinity for the ALK epitope characterized by a K d of 30 nM or less.
14. The isolated nucleic acid of claim 11 , said antibody or fragment having a binding affinity for the ALK epitope characterized by a K d of less than 3 nM.
15. The isolated nucleic acid of claim 11 , said antibody comprising the structure NH 2 —VL-linker-VH—COOH or NH 2 —VH-linker-VL-COOH, wherein the linker has the sequence GGGGSGGGGSGGGGSSGGGS (SEQ ID NO: 16).