Biological materials and uses thereof

• US 20060193849A1
• Filed: 02/25/2005
• Published: 08/31/2006
• Est. Priority Date: 02/25/2005
• Status: Abandoned Application

• Current Assignee: Antisoma PLC (Sarossa Plc), The United States of America As Represented By The Secretary of Agriculture
• Original Assignee: National Institutes of Health (NIH) (Government of the United States of America), Antisoma PLC (Sarossa Plc)
• Inventors: Krauss, Juergen, Courtenay-Luck, Nigel S., Rybak, Susanna M.
• Application Number:
  US11/066,045
• Publication Number:
  US 20060193849A1
• Time in Patent Office: Days
• US Class Current: 424/133.1
• CPC Class Codes: A61K 47/6815 Enzymes
• CPC Class Codes: A61K 47/6851 the antibody targeting a determinant of a tumour cell
• CPC Class Codes: C07K 16/3092 against tumour-associated mucins
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value

Abstract

A modified antibody molecule which selectively binds to a specific target, the antibody molecule being modified at at least one amino acid residue that determines antigen binding selectivity and/or affinity, characterised in that the modified antibody molecule exhibits a greater stability than an unmodified parent antibody molecule which selectively binds to the target. Also described are nucleotide sequences encoding, vectors, host cells and composition containing and uses of such antibodies, antibody fragments or antibody derivatives.

Claims

1. A modified antibody molecule which selectively binds to a specific target, the antibody molecule being modified at, at least one amino acid residue that determines antigen binding selectivity and/or affinity, wherein the modified antibody molecule exhibits a greater stability than an unmodified parent antibody molecule, which selectively binds to that target.

2. The modified antibody molecule of claim 1, wherein the site of modification is at one or more of the corners of the inner faces of the beta-sheets of the antibody molecule.

3. The modified antibody molecule of claim 1, wherein the site of the modification is the amino acid residue V_H71 in the amino acid sequence of FIG. 8 (SEQ ID NO:

1) or the corresponding residue in another antibody molecule.

4. The modified antibody molecule of claim 1, wherein the specific target is the MUC-1 gene product.

5. The modified antibody molecule of claim 1, wherein the modified amino acid possesses at least one physicochemical property different to the amino acid before modification.

6. The modified antibody molecule of claim 5, wherein the physicochemical property is at least one selected from group consisting of charge, hydrophobicity/hydrophilicity, size, structure, volume, polarity, and side chain characteristics.

7. The modified antibody molecule of claim 6, wherein the size of the amino acid is reduced by the modification.

8. The modified antibody molecule of claim 1, wherein the modification results in the amino acid, alanine.

9. The modified antibody molecule of claim 1, wherein the modification increases the buried surface area between motifs of the antigen binding site.

10. The modified antibody molecule of claim 1, wherein the modified antibody molecule has an antigen binding selectivity equivalent to gamma 1, kappa anti-HMFG monoclonal antibody.

11. The modified antibody molecule of claim 1, wherein the modified antibody molecule is a single chain antibody.

12. The modified antibody molecule of claim 11, wherein the single chain antibody is a single chain Fv.

13. The modified antibody molecule of claim 12, wherein the single chain antibody is a diabody.

14. The modified antibody molecule of claim 1, wherein the modified antibody molecule is humanized.

15. The modified antibody molecule of claim 14, wherein a humanized amino acid residue is modified to the murine amino acid.

16. The modified antibody molecule of claim 1, wherein the modified antibody molecule is associated with at least one other agent.

17. The modified antibody molecule of claim 16, wherein the agent is selected from the group consisting of drugs, toxins, radionuclides, nucleases, enzymes, cytokines and chemokines.

18. The modified antibody molecule of claim 16, wherein the agent is conjugated to the modified antibody molecule.

19. The modified antibody of claim 16, wherein the agent is a nuclease.

36. The nucleotide sequence of claim 5 wherein the nucleotide sequence is operably linked to the nucleotide sequence encoding a phage surface protein.

37. The nucleotide sequence of claim 6 wherein the phage expresses and displays the product of the sequence.

20. A nucleotide sequence encoding a modified antibody molecule, which selectively binds to a specific target, the antibody molecule being modified at, at least one amino acid residue that determines antigen binding selectivity and/or affinity, wherein the modified antibody molecule exhibits a greater stability than an unmodified parent antibody molecule, which selectively binds to that target.

21. The nucleotide sequence of claim 20, wherein the nucleotide sequence is that of FIG. 8 (SEQ ID NO:

2).

22. An expression vector containing a nucleotide sequence encoding a modified antibody molecule, which selectively binds to a specific target, the antibody molecule being modified at, at least one amino acid residue that determines antigen binding selectivity and/or affinity, wherein the modified antibody molecule exhibits a greater stability than an unmodified parent antibody molecule, which selectively binds to that target.

23. The expression vector of claim 22, wherein the nucleotide sequence is that of FIG. 8 (SEQ ID NO:

2).

35. The nucleotide sequence of claim 22 wherein the sequence is in an expression phage.

38. A method of screening of antibodies, antibody fragments or antibody derivatives that are able to bind a target molecule comprising providing the phage of claim 35 and one or more target molecules.

39. The method of claim 38 wherein the target molecules is a MUC1 gene product.

24. A host cell producing a modified antibody molecule, which selectively binds to a specific target, the antibody molecule being modified at, at least one amino acid residue that determines antigen binding selectivity and/or affinity, wherein the modified antibody molecule exhibits a greater stability than an unmodified parent antibody molecule, which selectively binds to that target, resulting from the expression of the nucleotide sequence encoding the modified antibody molecule.

25. The host cell of claim 24, wherein the nucleotide sequence is that of FIG. (SEQ ID NO:

2).

26. A composition comprising a modified antibody molecule, which selectively binds to a specific target, the antibody molecule being modified at, at least one amino acid residue that determines antigen binding selectivity and/or affinity, wherein the modified antibody molecule exhibits a greater stability than an unmodified parent antibody molecule, which selectively binds to that target, and a pharmaceutically acceptable carrier, excipient and/or diluent.

28. The composition of claim 26 further comprising at least one other agent.

30. The composition of claim 28 wherein, the agent is selected from the group consisting of drugs, toxins, radionuclides, nucleases, enzymes, cytokines and chemokines.

27. A composition comprising a nucleotide sequence encoding a modified antibody molecule, which selectively binds to a specific target, the antibody molecule being modified at, at least one amino acid residue that determines antigen binding selectivity and/or affinity, wherein the modified antibody molecule exhibits a greater stability than an unmodified parent antibody molecule, which selectively binds to that target.

29. The composition of claim 27 further comprising at least one other agent.

31. The composition of claim 29 wherein, the agent is selected from the group consisting of drugs, toxins, radionuclides, nucleases, enzymes, cytokines and chemokines.

32. A method for the treatment and/or diagnosis and/or prevention of a disorder selected from the group consisting of cancer, inflammatory disorders, cardiovascular diseases, infectious diseases, autoimmune disorders, central nervous system disorders, nephritis, sepsis, haemoglobinuria, chemotherapy induced thrombocytopenia, and addiction comprising administering an effective amount of a modified antibody molecule, which selectively binds to a specific target, the antibody molecule being modified at, at least one amino acid residue that determines antigen binding selectivity and/or affinity, wherein the modified antibody molecule exhibits a greater stability than an unmodified parent antibody molecule, which selectively binds to that target.

33. The method of claim 32, wherein the disease is cancer.

34. The method of claim 33, wherein the cancer is selected from the group consisting of cancer of the breast, ovary, uterus, lung, B-cell non-Hodgkins lymphoma, multiple myeloma, acute myelogenous leukemia, chronic lymphocytic leukemia and hairy cell leukemia.