Polynucleotides encoding anti-GD2 antibodies
Described herein are antibodies that specifically bind ganglioside GD2. Also described are nucleotides encoding such antibodies, cells expressing such antibodies, methods of use for such antibodies, and methods for using the antibodies to treat diseases associated with ganglioside GD2. In addition, tissue culture media supplements are described as are methods of use for the supplements. Described herein are albumin-ganglioside conjugates and corresponding methods for producing such conjugates. Methods of purifying or isolating antibodies are also described.
1. An isolated polynucleotide encoding an antibody, or antigen binding fragment thereof, that specifically binds to human GD2, having a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 11, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 12, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 26, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 28.
2. The isolated polynucleotide of claim 1 , comprising the nucleic acid sequences of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 18, SEQ ID NO: 19, and SEQ ID NO: 20.
3. The isolated polynucleotide of claim 1 , comprising the nucleic acid sequences of SEQ ID NO: 8 and SEQ ID NO: 24.
4. The isolated polynucleotide of claim 1 , comprising the nucleic acid sequences of SEQ ID NO: 39 and SEQ ID NO: 41.
5. A vector comprising the isolated polynucleotide of claim 1 .
6. An isolated cell comprising the vector of claim 5 .
7. The isolated cell of claim 6 , wherein the cell is a bacterial cell.
8. The isolated cell of claim 6 , wherein the cell is a eukaryotic cell.
9. The isolated eukaryotic cell of claim 8 , wherein the cell is a Chinese Hamster Ovary (CHO) cell.
10. The isolated eukaryotic cell of claim 8 , wherein the cell does not express J-chain.
11. The isolated eukaryotic cell of claim 10 , wherein the cell produces a pentameric IgM antibody.
12. The isolated eukaryotic cell of claim 10 , wherein the cell produces more pentameric IgM than hexameric IgM.
13. A method of making an antibody or antigen-binding fragment thereof that specifically binds to human GD2 and includes a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 11, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 12, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 26, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 28, comprising culturing the isolated eukaryotic cell of claim 10 under conditions suitable to produce the antibody or antigen-binding fragment thereof and recovering the antibody or antigen-binding fragment thereof from the cell culture.
14. A method of making an antibody or antigen-binding fragment thereof that specifically binds to human GD2 and includes a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 11, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 12, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 26, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 28, comprising culturing the isolated cell of claim 6 under conditions suitable to produce the antibody or antigen-binding fragment thereof and recovering the antibody or antigen-binding fragment thereof from the cell culture.
15. A recombinant polynucleotide encoding an antibody heavy chain, or a fragment thereof, of an antibody that specifically binds to human GD2, wherein the heavy chain CDR1 of the antibody comprises the amino acid sequence of SEQ ID NO: 10, the heavy chain CDR2 of the antibody comprises the amino acid sequence of SEQ ID NO: 11, and the heavy chain CDR3 of the antibody comprises the amino acid sequence of SEQ ID NO: 12, wherein the encoded heavy chain or fragment thereof comprises SEQ ID NOs: 10, 11, and 12.
16. The recombinant polynucleotide of claim 15 , comprising the nucleic acid sequences of SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4.
17. The recombinant polynucleotide of claim 15 , comprising the nucleic acid sequence of SEQ ID NO: 8.
18. The recombinant polynucleotide of claim 15 , comprising the nucleic acid sequence of SEQ ID NO: 39.
19. A vector comprising the recombinant polynucleotide of claim 15 .
20. An isolated cell comprising the vector of claim 19 .
21. The isolated cell of claim 20 , wherein the cell is a bacterial cell.
22. The isolated cell of claim 20 , wherein the cell is a eukaryotic cell.
23. The isolated eukaryotic cell of claim 22 , wherein the cell is a Chinese Hamster Ovary (CHO) cell.
24. An isolated cell comprising a vector having a recombinant polynucleotide encoding an antibody heavy chain, or a fragment thereof, of an antibody that specifically binds to human GD2, wherein the encoded heavy chain or fragment thereof includes a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO:10, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO:11, and a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO:12, the isolated cell further comprising a separate vector having a recombinant polynucleotide encoding an antibody light chain, or a fragment thereof, of an antibody that specifically binds to human GD2, wherein the encoded light chain or fragment thereof includes a light chain CDR1 comprising the amino acid sequence of SEQ ID NO:26, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO:27, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO:28.
25. A method of making an antibody or antigen-binding fragment thereof that specifically binds to human GD2 and includes a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 10, a heavy chain CDR2 comprising the amino acid sequence of SEQ ID NO: 11, a heavy chain CDR3 comprising the amino acid sequence of SEQ ID NO: 12, a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 26, a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 27, and a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 28, comprising culturing the isolated cell of claim 24 under conditions suitable to produce the antibody or antigen-binding fragment thereof and recovering the antibody or antigen-binding fragment thereof from the cell culture.
26. The method of claim 25 , wherein the isolated cell is a eukaryotic cell that does not express J-chain.
27. The method of claim 26 , wherein the antibody is a recombinant pentameric IgM antibody.
28. The method of claim 26 , wherein the isolated cell produces more pentameric IgM than hexameric IgM.