Tumour marker proteins and uses thereof

Abstract

Tumour marker proteins and their preparation from fluids from one or more cancer patients, wherein said fluids are those which collect in a body cavity or space which is naturally occurring or which is the result of cancer or medical intervention for cancer. The present application also relates to preparation of tumour marker proteins from excretions taken from patients with cancer. The tumour marker proteins are useful as immunoassay reagents in the detection of cancer-associated anti-tumour marker autoantibodies.

What is claimed: 
     
         1 . A method of detecting cancer-associated anti-tumor autoantibodies in a sample from an individual, comprising:
 contacting the sample with an immunoassay reagent; and detecting a presence of complexes formed by specific binding of the immunoassay reagent to any cancer-associated anti-tumor autoantibodies present in the sample,   wherein the immunoassay reagent comprises two or more tumor marker proteins, one or more of which having been prepared from a tumor-induced bodily fluid produced in a body cavity or space in the presence of the tumor of one or more cancer patients, wherein the bodily fluid contains more cancer-associated forms of the tumor marker protein than a non-tumor-induced bodily fluid in the same patient, and the bodily fluid is not a fluid from the systemic circulation,   wherein the one or more tumor marker proteins prepared from a bodily fluid exhibit selective reactivity with cancer-associated anti-tumor autoantibodies, wherein the tumor marker proteins are over-expressed or altered forms of wild-type proteins, and   wherein detection of complexes indicates the presence of cancer-associated anti-tumor autoantibodies in the individual.   
     
     
         2 . The method of  claim 1 , further comprising detecting and/or quantitatively measuring the presence of two or more types of autoantibodies, wherein each one of the two or more types of the autoantibodies is immunologically specific to a different tumor marker protein or to different epitopes of the same tumor marker protein, wherein the immunoassay is carried out using a panel of the two or more tumor marker proteins. 
     
     
         3 . The method of  claim 2 , wherein the immunoassay is carried out using a panel of two or more immunoassay reagents, wherein each one of the two or more immunoassay reagents is for detection of a different tumor marker protein, wherein the sample is obtained from a patient, the method further comprising:
 determining relative strength of immune response of the patient to each one of the two or more different tumor marker proteins; and   selecting one or more tumor marker proteins to form a basis of an anti-cancer vaccine for use in the patient, wherein the one or more tumor marker proteins to which the patient has strongest immune response is selected out of the two or more different tumor marker proteins.   
     
     
         4 . The method of  claim 1 , wherein the bodily fluid is ascites fluid, pleural effusion, seroma, hydrocoele or wound drainage fluid. 
     
     
         5 . The method of  claim 1 , wherein the excretion is urine, faeces or seminal fluid. 
     
     
         6 . The method of  claim 1 , wherein the one or more tumor marker proteins prepared from the bodily fluid is MUC1, MUC16, c-myc, c-erbB2, p53, ras, BRCA1, BRCA2, APC, PSA, CEA, or CA19.9. 
     
     
         7 . A method of determining whether a vaccination procedure, comprising challenging a patient with an immunogenic preparation comprising a tumor marker protein or an antigenic fragment thereof or with a nucleic acid sequence expressing the tumor marker protein, has been successful in eliciting cancer-associated antibodies to the tumor marker protein in the patient, wherein the method is an immunoassay, comprising:
 contacting a sample of bodily fluid from the patient with an immunoassay reagent; and   detecting presence of complexes formed by specific binding of the immunoassay reagent to any cancer-associated antibodies present in the sample,   wherein the immunoassay reagent comprises a sample of the tumor marker protein,   wherein the tumor marker protein is prepared from a bodily fluid derived from one or more of a body cavity or in which a tumor is or was present or with which a tumour is or was associated, of one or more cancer patients, or from an excretion from one or more cancer patients, wherein the tumor marker protein exhibits selective reactivity with cancer-associated anti-tumor antibodies.   
     
     
         8 . A method of preparing a tumour marker protein wherein the method comprises isolating the tumour marker protein from bodily fluid wherein the fluid is: (i) collected from a body cavity or space in which a tumour is or was present or with which a tumour is or was associated, and (ii) the fluid represents the pooled fluid samples from two or more cancer patients. 
     
     
         9 . The method of  claim 8  wherein the fluid is ascites, pleural effusion, seroma, hydrocoele or wound drainage fluid or a mixture thereof. 
     
     
         10 . The method of  claim 8  wherein the tumour marker protein is MUC1, c-erbB2, p53, ras, BRCA1, BRCA2, APC, PSA, CEA, CA19.9, MUC16 or c-myc. 
     
     
         11 . A method of preparing a tumour marker protein wherein the method comprises isolating the tumour marker protein from a bodily fluid collected from a body cavity or space in which a tumour is or was present or with which a tumour is or was associated, and wherein the bodily fluid is wound drainage fluid, seroma, hydrocoele or a mixture thereof. 
     
     
         12 . A method of preparing a tumour marker protein wherein the method comprises isolating the tumour marker protein from an excretion wherein: (i) the excretion or any component thereof has been in contact with a tumour or tumour cells, and (ii) the excretion represents pooled excretion samples from two or more cancer patients. 
     
     
         13 . The method of  claim 12  wherein the excretion is urine, faeces or seminal fluid. 
     
     
         14 . The method of  claim 12  wherein the relevant component of the excretion is bile. 
     
     
         15 . The method of  claim 12 , wherein the tumour marker protein is MUC1, c-erbB2, p53, ras, BRCA1, BRCA2, APC, PSA, CEA, CA19.9, MUC16 or c-myc. 
     
     
         16 . The method of  claim 12  wherein the tumour marker is purified from the fluid or excretion by affinity chromatography. 
     
     
         17 . The method of  claim 12 , further comprising removing contaminating immunoglobulin from the tumour marker protein. 
     
     
         18 . The method of  claim 12 , further comprising immobilizing the isolated tumour marker protein on a solid support. 
     
     
         19 . A tumour marker protein prepared by the method of  claim 12 , wherein the tumour marker protein is substantially immunoglobulin free. 
     
     
         20 . A kit or reagent suitable for carrying out an immunoassay, wherein the kit or reagent comprises the tumour marker protein of  claim 19  immobilized on a solid support. 
     
     
         21 . The kit or reagent of  claim 20 , wherein the solid support is the surface of a well of a multiwell plate or a bead. 
     
     
         22 . The kit or reagent of  claim 20 , wherein the immobilized tumour marker protein is absorbed, adsorbed or covalently attached to the solid support. 
     
     
         23 . A method of calibrating an assay for measurement or detection of a tumour marker protein in a clinical sample, wherein the method comprises the steps of:
 a) preparing at least two samples of the tumour marker protein of  claim 19 , each of which comprises the given tumour marker protein and each of which has a different tumour marker protein concentration to each of the other the samples:   b) carrying out a quantitative measurement of the concentration of the tumour marker protein in each of the samples using (i) a spectrophotometric method and/or, (ii) an antibody reagent to the tumour marker protein, and   c) constructing a standard curve for a tumour marker protein concentration based on the measurements obtained in step (b).