Antibodies specific for CYP1B1

Abstract

Antibodies that can specially bind to cytochrome P450 CYP1B1 and methods of making them are disclosed, in particular antibodies that bind to amino acid sequence VNQWSVNHDPVKWPN or PExFDPARFLDKDGy, where x is D or N and y is L or F, or an antigenic fragment thereof. The antibodies can be used in the diagnosis or treatment of cancers linked to enhanced CYP1B1 expression, including breast cancer, prostate cancer, colorectal cancer, liver cancer and ovarian cancer.

1 - 25 . (canceled) 
     
     
         26 . An assay method for detecting cancer cells present in a sample from a patient, the method comprising:
 contacting a tissue sample from the patient with an antibody which is capable of specifically binding to cytochrome P450 CYP1B1, wherein the antibody recognizes an epitope in the cytochrome P450 CYP1B1 protein included within the amino acid sequence VNQWSVNHDPVKWPN (SEQ ID NO:1) or PExFDPARFLDKDGy, where x is D or N and y is L or F (SEQ ID NOs: 2 to 5); and,   detecting binding of the antibody to CYP1B1 protein present in the sample as an indication of the presence of cancer cells in the tissue sample.   
     
     
         27 . The method of  claim 26 , wherein the antibody recognizes an epitope of between 3 and 10 amino acids from the amino acid sequence VNQWSVNHDPVKWPN (SEQ ID NO: 1) or PExFDPARFLDKDGy, where x is D or N and y is L or F (SEQ ID NOs: 2 to 5). 
     
     
         28 . The method of  claim 26 , wherein the antibody recognizes an epitope of between 3 and 6 amino acids from the amino acid sequence VNQWSVNHDPVKWPN (SEQ ID NO: 1) or PExFDPARFLDKDGy, where x is D or N and y is L or F (SEQ ID NOs: 2 to 5). 
     
     
         29 . The method of  claim 26 , wherein the antibody is a monoclonal antibody. 
     
     
         30 . The method of  claim 26 , wherein the antibody is humanized. 
     
     
         31 . The method of  claim 29 , wherein the antibody is a monoclonal antibody and is obtainable by:
 (a) immunizing an animal with a peptide comprising the amino acid sequence VNQWSVNHDPVKWPN (SEQ ID NO: 1) or PExFDPARFLDKDGy, where x is D or N and y is L or F (SEQ ID NOs: 2 to 5), which peptide is conjugated to an immunogenic carrier;   (b) sacrificing the animal and fusing spleen cells obtained from the animal with myeloma cells to produce one or more hybridomas; and   (c) screening the hybridomas for antibodies capable of binding the peptide.   
     
     
         32 . The method of  claim 26 , wherein the antibody is conjugated to an effector. 
     
     
         33 . The method of  claim 32 , wherein the effector is selected from the group consisting of: a label and an enzyme. 
     
     
         34 . A method according to  claim 33 , wherein the label is selected from the group consisting of: a radioactive label and a fluorescent label. 
     
     
         35 . A method according to  claim 33 , wherein the enzyme is horseradish peroxidase. 
     
     
         36 . The method of  claim 26 , wherein the step of detecting the binding of the antibody to CYPB1 protein is carried out using an assay selected from the group consisting of: an antibody capture assay, a two-antibody sandwich assay and an antigen capture assay. 
     
     
         37 . The method of  claim 36 , wherein the antibody specific for the CYP1B1 protein is immobilized on a solid support. 
     
     
         38 . The method of  claim 37 , wherein the antibody is immobilized on the solid support at defined, spatially separated locations. 
     
     
         39 . The method of  claim 26 , wherein the cancer is selected from the group consisting of: breast cancer, colorectal cancer, prostate cancer, liver cancer and ovarian cancer. 
     
     
         40 . The method of  claim 26 , wherein said tissue sample is selected from the group consisting of: bladder, brain, breast, colon, connective tissue, kidney, lung, lymph node, oesophagus, ovary, skin, stomach, testis, uterus, prostate and liver.