Preparation and application of human HCC-associated antigen Cdc25C specific peptide antibody
CHEN Chengxiao
ZHANG Qihui
LI Chunmei
TIAN Lin
MO Farong
Abstract：Objective To prepare polyclonal antibody against human hepatocellular carcinoma ( HCC)-associated an-tigen Cdc25C specific polypeptide, and to lay a foundation for further study of the function of Cdc25C.Methods The properties of Cdc25C protein antigen were analyzed by bioinformatics software.Several candidate partial peptides of low ho-mology and high antigenicity were designed based on the purity, amino acid composition, length, hydrophilic and secondary structure.Three partial peptides of Cdc25C were synthesized.The synthesized peptide was then used to immunize after cou-pling with KLH.The activity of anti-Cdc25C antibody was analyzed by ELISA and Western blotting, and its effect was de-tected by immunohistochemistry.Results The polypeptide antibody had high titer and specificity.The antibody identified against recombinant Cdc25C fusion protein by Western blotting, and the reaction bands appeared at 73 kD.We used the whole protein monoclonal antibody and peptide antibody to detect the same HCC cancer adjacent tissue, and its average op-tical density was 0.538 ±0.192 and 0.410 ±0.122, respectively, no significant difference was found between them ( t=0.109, P>0.05).Conclusion Polyclonal antibody against human HCC-associated antigen Cdc25C specific polypeptide is successfully prepared.The candidate peptides obtained from the screening have the same antigenicity as Cdc25C whole protein, and the prepared polypeptide antibody has the same effect as the antibody against the Cdc25C whole protein.
Keywords：liver neoplasmshepatocellular carcinoma-associated antigenCdc25C polypeptidepolyclonal antibody
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 1-3 )