Antibodies specific for claudin 6 (CLDN6)
The present invention provides antibodies useful as therapeutics for treating and/or preventing diseases associated with cells expressing Claudin-6 (CLDN6), including tumor-related diseases such as ovarian cancer, lung cancer, gastric cancer, breast cancer, hepatic cancer, pancreatic cancer, skin cancer, malignant melanoma, head and neck cancer, sarcoma, bile duct cancer, cancer of the urinary bladder, kidney cancer, colon cancer, placental choriocarcinoma, cervical cancer, testicular cancer, and uterine cancer.
1. A method of producing an anti-CLDN6 antibody that binds to CLDN6 or an antigen binding fragment thereof, the method comprising the steps of:
a. culturing a host cell transformed with one or more expression vectors encoding the antibody or antigen binding fragment thereof under conditions in which the host cell expresses the antibody or antigen binding fragment thereof; and
b. harvesting a preparation of the antibody or antigen binding fragment thereof expressed by the cell;
wherein the one or more expression vectors comprise:
(i) a nucleic acid sequence encoding heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 34 and a nucleic acid encoding light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 35,
(ii) a nucleic acid sequence encoding heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 36 and a nucleic acid encoding light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 37,
(iii) a nucleic acid sequence encoding heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 38 and a nucleic acid encoding light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 39, or
(iv) a nucleic acid sequence encoding heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 40 and a nucleic acid encoding light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 41.
2. The method of claim 1 , wherein said host cell is selected from the group consisting of a lymphocytic cell, eukaryotic host cell, CHO cell, NS/0 cell, HEK293 cell, HEK293T cell, plant cell, fungal cell, dendritic cell, B cell, COS cell, K562 cell, HELA cell, yeast cell, and insect cell, preferably a CHO cell.
3. The method of claim 1 , wherein said expression vector comprises a promoter sequence, a leader sequence, a translation initiation sequence, a light chain constant region, a heavy chain constant region, 3′ untranslated sequence, a polyadenylation sequence, or a transcription termination sequence.
4. A recombinant nucleic acid comprising a nucleic acid sequence selected from the group consisting of:
(i) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 34,
(ii) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 36,
(iii) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 38, and
(iv) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 40.
5. The recombinant nucleic acid of claim 4 , wherein said nucleic acid sequence encodes a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 34, 36, 38 and 40.
6. The recombinant nucleic acid of claim 4 , wherein the recombinant nucleic acid comprises a human or murine heavy chain constant region.
7. The recombinant nucleic acid of claim 6 , wherein said nucleic acid encoding said human heavy chain constant region comprises the nucleic acid sequence of SEQ ID NO: 24 or encodes the amino acid sequence of SEQ ID NO: 25.
8. A recombinant nucleic acid comprising a nucleic acid sequence selected from the group consisting of:
(i) a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 35,
(ii) a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 37,
(iii) a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 39, and
(iv) a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 41.
9. The recombinant nucleic acid of claim 8 , wherein said nucleic acid sequence encodes a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 35, 37, 39 and 41.
10. The recombinant nucleic acid of claim 8 , wherein the recombinant nucleic acid comprises a human or murine light chain constant region.
11. The recombinant nucleic acid of claim 10 , wherein said nucleic acid encoding said human light chain constant region comprises the nucleic acid sequence of SEQ ID NO: 26 or encodes the amino acid sequence of SEQ ID NO: 27.
12. The recombinant nucleic acid of claim 4 , wherein said nucleic acid sequence is operatively linked to expression control sequences.
13. The recombinant nucleic acid of claim 12 , wherein said expression control sequence allows expression in a prokaryotic or eukaryotic host cell.
14. The recombinant nucleic acid of claim 8 , wherein said nucleic acid sequence is operatively linked to expression control sequences.
15. The recombinant nucleic acid of claim 14 , wherein said expression control sequence allows expression in a prokaryotic or eukaryotic host cell.
16. A transformed cell comprising one or more expression vectors, the one or more expression vectors comprising:
(i) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 34 and a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 35,
(ii) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 36 and a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 37,
(iii) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 38 and a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 39, and
(iv) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 40 and a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 41.
17. The transformed cell of claim 16 , wherein said cell is selected from the group consisting of a lymphocytic cell, eukaryotic host cell, CHO cell, NS/0 cell, HEK293 cell, HEK293T cell, plant cell, fungal cell, dendritic cell, B cell, COS cell, K562 cell, HELA cell, yeast cell, and insect cell, preferably a CHO cell.
18. An anti-CLDN6 antibody or antigen binding fragment thereof, that binds to CLDN6, wherein said antibody, or antigen binding fragment thereof is obtained from the transformed cell of claim 16 .
19. A method of producing an antibody producing cell, comprising the steps of
a. transforming a cell with one or more expression vectors, the one or more expression vectors comprising:
(i) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 34 and a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 35,
(ii) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 36 and a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 37,
(iii) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 38 and a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 39, and
(iv) a nucleic acid sequence encoding a heavy chain of an antibody comprising heavy chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 40 and a nucleic acid sequence encoding a light chain of an antibody comprising light chain CDR1, CDR2, and CDR3 regions of SEQ ID NO: 41; and
b. obtaining the transformed cell, wherein the transformed cell comprises the nucleic acid sequence encoding a heavy chain of an antibody and the nucleic acid sequence encoding a light chain of an antibody.