Anti-p53 antibodies
The invention relates to methods for producing an antibody which is specific for a mutant p53 polypeptide over wildtype p53 polypeptide, comprising using the mutant p53 polypeptide as an immunogen wherein the polypeptide comprises: (i) an antigen sequence, comprising an amino acid sequence of the mutant p53 polypeptide including the mutation and at least one amino acid immediately adjacent to the mutation, and (ii) a scaffold sequence for providing the antigen sequence in a solvent-accessible configuration, in particular wherein the scaffold sequence is thioredoxin. In the specific embodiments, antibodies against mutant p53 comprising R175H, R248Q or R273H are generated. Also disclosed are the uses of the antibodies for diagnosis, prognosis and stratification of patient groups and further encompasses the use of antibodies for imaging and treatment of cancer.
1. A method for producing an antibody which is specific for a mutant p53 polypeptide over wildtype p53 polypeptide, comprising administering to a subject an immunogen, wherein the immunogen is a peptide or polypeptide comprising: (i) an antigen sequence, comprising at least two amino acid sequences of the mutant p53 polypeptide including the mutation and at least one amino acid immediately adjacent to the mutation, wherein the at least two amino acid sequences of the mutant p53 polypeptide are non-identical; and (ii) a scaffold sequence derived from a thioredoxin for providing the antigen sequence in a solvent-accessible configuration.
2. The method according to claim 1 , wherein the antigen sequence is inserted in, or substituted for all or part of, the active site sequence of the thioredoxin.
3. The method according to claim 1 , wherein the peptide or polypeptide used as an immunogen additionally comprises one or more linker sequences between the antigen sequence and the scaffold sequence.
4. The method according to claim 1 , wherein the peptide or polypeptide used as an immunogen additionally comprises linker sequences between the at least two amino acid sequences of the mutant p53 polypeptide.
5. The method according to claim 1 , wherein the amino acid sequence of the mutant p53 polypeptide comprises at least 5 amino acids.
6. The method according to claim 1 , wherein the mutant p53 polypeptide comprises a mutation in the DNA-binding domain (DBD).
7. The method according to claim 1 , wherein the mutant p53 polypeptide comprises a mutation selected from one of R175H, R248Q, R273H, R248W, G245S, R273C, R282W, R249S, G245D, C176F, H179Y, H179R, Y220C and R337H.
8. The method according to claim 1 , wherein the mutant p53 polypeptide comprises, or consists of, the amino acid sequence of one of SEQ ID NOs: 3 to 16.
9. A cancer vaccine, comprising an immunogen, wherein the immunogen is a peptide or polypeptide comprising: (i) an antigen sequence, comprising at least two amino acid sequences of a mutant p53 polypeptide including the mutation and at least one amino acid immediately adjacent to the mutation, wherein the at least two amino acid sequences of the mutant p53 polypeptide are non-identical; and (ii) a scaffold sequence derived from a thioredoxin for providing the antigen sequence in a solvent-accessible configuration.
10. The cancer vaccine of claim 9 , wherein the cancer is a cancer comprising a cell or cells expressing a mutant p53 polypeptide, or comprising nucleic acid encoding a mutant p53 polypeptide.
11. A method for vaccinating a subject against a cancer, comprising administering to the subject the cancer vaccine of claim 9 , thereby vaccinating the subject against a cancer.
12. The method of claim 11 , wherein the cancer is a cancer comprising a cell or cells expressing a mutant p53 polypeptide, or comprising nucleic acid encoding a mutant p53.