Claims
- 1. A mutant antibody comprising a reactive site not present in the wildtype of said antibody and an antigen recognition domain that recognizes a macrocyclic metal chelate comprising four nitrogen atoms, wherein said reactive site is in a position proximate to or within said antigen recognition domain.
- 2. The mutant antibody of claim 1, wherein at least two of said nitrogen atoms are covalently linked to a substituted or unsubstituted ethyl bridge.
- 3. The mutant antibody of claim 2, wherein said metal chelate comprises the subunit:
- 4. The mutant antibody of claim 1, wherein said chelate is a member selected from substituted or unsubstituted DOTA and substituted or unsubstituted TETA.
- 5. The mutant antibody of claim 4, wherein said chelate has the formula:
- 6. The mutant antibody of claim 1, wherein said macrocyclic metal chelate comprises a reactive functional group having reactivity complementary to said reactive site of said mutant antibody.
- 7. The mutant antibody of claim 5, wherein the carbon atom marked is of S configuration.
- 8. The mutant antibody of claim 1, comprising a moiety having the formula:
- 9. The mutant antibody according to claim 1, wherein said reactive site is a side-chain of a naturally occurring or non-naturally occurring amino acid.
- 10. The mutant antibody according to claim 9, wherein said reactive site is the —SH group of cysteine.
- 11. The mutant antibody according to claim 9, further comprising said metal chelate bound to said antigen recognition domain, wherein said metal chelate comprises a reactive functional group of complementary reactivity to said reactive site of said antibody.
- 12. The mutant antibody according to claim 11, further comprising a covalent bond between reactive site of said antibody and said reactive functional group of said metal chelate.
- 13. The mutant antibody according to claim 1, wherein said mutant antibody is mutant of 2D12.5.
- 14. The mutant antibody according to claim 13, wherein asparagine-88 residue of the heavy-chain is substituted by an aspartic acid residue.
- 15. The mutant antibody according to claim 14, wherein glycine-54 residue of the heavy-chain is substituted by a cysteine residue.
- 16. The mutant antibody according to claim 14, wherein glycine-55 residue of the heavy-chain is substituted by a cysteine residue.
- 17. The mutant antibody according to claim 14, wherein glycine-56 residue of the heavy-chain is substituted by a cysteine residue
- 18. The mutant antibody according to claim 13, wherein asparagine-54 of the light chain is substituted by a cysteine residue.
- 19. The mutant antibody of claim 1, further comprising a targeting moiety.
- 20. The mutant antibody according to claim 19, wherein said targeting moiety binds specifically to a cell surface protein.
- 21. The mutant antibody according to claim 19, wherein the targeting moiety is covalently attached to said mutant antibody.
- 22. The mutant antibody according to claim 20, wherein the targeting moiety is an antibody.
- 23. The mutant antibody according to claim 21, wherein the targeting moiety specifically binds to a protein on a cancer cell.
- 24. A composition having the structure:
- 25. The composition of claim 24, wherein said targeting moiety is an antibody that binds specifically to a cell surface antigen.
- 26. A pharmaceutical composition comprising the composition of claim 24, and a pharmaceutically acceptable carrier.
- 27. A method of in vivo imaging, said method comprising the steps of:
(a) administering a pretargeting reagent to a subject; (b) administering to said subject a mutant antibody of claim 1, wherein said antibody comprises a recognition moiety that binds specifically to said pretargeting reagent, thereby forming a pretargeting reagent-mutant antibody complex; (c) administering to said subject said metal chelate, thereby specifically binding said compound to said mutant antibody to form a pretargeting reagent-mutant antibody-metal chelate complex; and (d) detecting said mutant antibody-metal chelate complex.
- 28. The method of claim 27, wherein the step of detecting is by positron emission tomography.
- 29. The method of claim 27, wherein the step of detecting is by magnetic resonance imaging.
- 30. The method of claim 27, wherein the step of detecting is by detection of lanthanide luminescence.
- 31. The method of claim 27, further comprising, between steps (a) and (b), administering a clearing agent to said subject.
- 32. The method of claim 27, wherein the subject is a mammal.
- 33. The method of claim 32, wherein the mammal is a human.
- 34. A method of treating a subject with cancer by administration of a metal chelate, said method comprising the steps of:
(a) administering to said subject a mutant antibody of claim 1, wherein said antibody comprises a targeting moiety that binds specifically to a cell by binding with a member selected from the group consisting of cell surface receptors and cell surface antigens, thereby forming a cell-mutant antibody complex; and (b) administering to said subject said metal chelate, thereby specifically binding said compound to mutant antibody to form a cell-mutant antibody-metal chelate complex.
- 35. The method of claim 34, wherein the subject is a mammal.
- 36. The method of claim 35, wherein the mammal is a human.
- 37. An isolated nucleic acid, comprising a sequence selected from a sequence as set forth in SEQ ID NOS:16, 17, 18, 19, 20, 25, 26, 31, 32, 42, 43, 44, 45, 46, and 47.
- 38. A polypeptide comprising an amino acid sequence selected from a sequence as set forth in SEQ ID NOS: 10, 11, 12, 13, 14, 22, 23, 27, 28, 35, 36, 37, 38, 39, and 40.
- 39. The isolated nucleic acid according to claim 37, further comprising a promoter operably linked to the nucleic acid sequence.
- 40. An expression vector comprising the nucleic acid according to claim 39.
- 41. A host cell comprising the expression vector according to claim 40.
STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT
[0001] This invention was made with Government support under Grant No. CA16861, awarded by the NIH/NCI to C. F. Meares. The Government has certain rights in this invention.