DETECTION OF ANTI-p53 ANTIBODIES

Abstract

The disclosure relates to an in vitro method for detecting an antibody to p53 (anti-p53 antibody) in a sample, the method comprising: incubating a sample to be analyzed with a p53 capture antigen and a p53 detection antigen, whereby a complex comprising the p53 capture antigen, the anti-p53 antibody and the p53 detection antigen is formed, separating the complex formed from unbound detection antigen and measuring the complex obtained via the detection antigen comprised therein, thereby detecting the anti-p53 antibody comprised in the sample.

1 . An in vitro method for detecting an antibody to p53 (anti-p53 antibody) in a sample, the method comprising:
 a) incubating a sample to be analyzed with a p53 capture antigen and a p53 detection antigen, whereby a complex comprising the p53 capture antigen the anti-p53 antibody and the p53 detection antigen is formed,   b) separating the complex formed in (a) from unbound detection antigen,   c) measuring the complex obtained in step (b) via the detection antigen comprised therein, thereby detecting the anti-p53 antibody comprised in the sample.   
     
     
         2 . The method according to  claim 1 ,
 wherein the p53 capture antigen is capable of binding to the solid phase.   
     
     
         3 . The method according to  claim 1 ,
 wherein the p53 capture antigen is covalently bound to a first partner of a binding pair and wherein the second partner of said binding pair is bound to the solid phase.   
     
     
         4 . The method according to  claim 1 ,
 wherein the first partner of the binding pair is selected from hapten, biotin or biotin analogues such as aminobiotin, iminobiotin or desthiobiotin, DsF and the ligand for a receptor and wherein the second partner of said binding pair is selected from anti-hapten antibody, avidin or streptavidin, FimG DsF, and a ligand-receptor.   
     
     
         5 . The method according to  claim 1 ,
 wherein the p53 capture antigen is biotinylated and wherein the solid phase is coated with avidin or streptavidin.   
     
     
         6 . The method according to  claim 1 ,
 wherein the p53 capture antigen is bound to the solid phase.   
     
     
         7 . The method according to  claim 1 ,
 wherein the p53 detection antigen comprises a directly detectable label.   
     
     
         8 . The method according to  claim 1 ,
 wherein the directly detectable label is selected from the group consisting of a luminescent label, a fluorescent label, a chemiluminescent label or an electrochemiluminescent label.   
     
     
         9 . The method according to  claim 1 ,
 wherein the directly detectable label is a chemiluminescent or an electrochemiluminescent label.   
     
     
         10 . The method according to  claim 1 ,
 wherein both the p53 capture antigen and the p53 detection antigen comprise a peptide of SEQ ID NO: 1.   
     
     
         11 . The method according to  claim 1 ,
 wherein both the p53 capture antigen and the p53 detection antigen comprise a peptide of SEQ ID NO: 2.   
     
     
         12 . The method according to  claim 1 ,
 wherein the peptides of SEQ ID NO:1 and of SEQ ID NO:2 are each comprised in both the p53 capture antigen and the p53 detection antigen.   
     
     
         13 . The method according to  claim 1 ,
 wherein the p53 detection antigen comprises both the peptides of SEQ ID NO:1 and of SEQ ID NO:2 at least twice.   
     
     
         14 . The method according to  claim 1 ,
 wherein the detection antigen comprises a fusion polypeptide comprising at least one multimerization domain, at least one polypeptide of SEQ ID NO: 1 and at least one polypeptide of SEQ ID NO:2, wherein the multimerization domain is a prokaryotic or eukaryotic chaperone selected from the group consisting of FkpA, Skp, SecB, Hsp25, MIP, GroEL, ClpB and ClpX.   
     
     
         15 . The method according to  claim 1 ,
 wherein the p53 detection antigen comprises an indirectly detectable label.   
     
     
         16 . A fusion polypeptide comprising at least one multimerization domain at least one polypeptide of SEQ ID NO: 1 and at least one polypeptide of SEQ ID NO:2, wherein the multimerization domain is a prokaryotic or eukaryotic chaperone selected from the group consisting of FkpA, Skp, SecB, Hsp25, MIP, GroEL, ClpB and ClpX.