山东医药2016，Vol.56Issue(13)：1-3,3.DOI:10.3969/j.issn.1002-266X.2016.13.001
人肝癌相关抗原 Cdc25C 特异性多肽抗体的制备及其应用
Preparation and application of human HCC-associated antigen Cdc25C specific peptide antibody
摘要
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.关键词
肝肿瘤/肝癌相关抗原/Cdc25C多肽/多克隆抗体Key words
liver neoplasms/hepatocellular carcinoma-associated antigen/Cdc25C polypeptide/polyclonal antibody分类
医药卫生引用本文复制引用
陈承晓,张琪惠,李春梅,田淋,莫发荣..人肝癌相关抗原 Cdc25C 特异性多肽抗体的制备及其应用[J].山东医药,2016,56(13):1-3,3.基金项目
国家自然科学基金资助项目（81160264）。 （）
