Synthetic peptide, relative artificial antigen, relative anti-ehd2 antibody and preparation method thereof and use thereof

Abstract

The present application provides a synthetic peptide with an amino acid sequence SEQ ID NO: 1 plus a cysteine at its N-terminal, an artificial antigen, an antibody specific to a human EHD2, a method for preparing an antibody specific to the human EHD2 protein and adopted to an immunohistochemical method for EHD2 detection as well as cancer diagnosing and prognosing.

1 . An antibody specific to a human Eps-15 homology domain-containing protein 2 (EHD2), wherein the antibody is obtained by
 administering to an animal an artificial antigen to obtain an antiserum, wherein the artificial antigen is a synthetic peptide coupling an antigen carrier, and the synthetic peptide consists of the amino acid sequence of SEQ ID NO: 1 with a cysteine added at the N-terminal end; and   performing antigen affinity purification of the antiserum to isolate said antibody.   
     
     
         2 . The antibody according to  claim 1 , wherein the antibody is a polyclonal antibody. 
     
     
         3 . The antibody according to  claim 1 , wherein the antibody is chemically labeled to add various enzymes or chromogenic groups, or various fluorophores, for direct detection of EHD2. 
     
     
         4 . The antibody according to  claim 1 , wherein the antibody is chemically labeled to add alkaline phosphatase, or various fluorophores, for direct detection of EHD2. 
     
     
         5 . A method of diagnosing or prognosing cancer, comprising immunohistochemical detecting of Eps-15 homology domain-containing protein 2 (EHD2) in human tissue samples by using the antibody of  claim 1  and by nuclear localization and quantification. 
     
     
         6 . The method according to  claim 5 , wherein the cancer is a breast cancer. 
     
     
         7 . The method according to  claim 5 , wherein the level of EHD2 expression in nuclei is negatively correlated with a malignancy degree of a tumor, and is positively correlated with a survival condition of a patient. 
     
     
         8 . The method according to  claim 5 , wherein said detecting of EHD2 in human tissue samples comprises using the antibody as a primary antibody to recognize EHD2, and using a secondary antibody or other reagents to conduct the subcellular EHD2 detection including nuclear and cytoplasm localization and quantification. 
     
     
         9 . The method according to  claim 5 , wherein said detecting of EHD2 in human tissue samples comprises labeling the antibody by adding various enzymes or chromogenic groups, or various fluorophores, and then directly using the labeled antibody for EHD2 detection without requiring a secondary antibody or other reagents. 
     
     
         10 . The method according to  claim 5 , wherein said detecting of EHD2 in human tissue samples comprises labeling the antibody by adding alkaline phosphatase, or various fluorophores, and then directly using the labeled antibody for EHD2 detection without requiring a secondary antibody or other reagents.