摘要
目的制备抗HCC-22-5的多克隆抗体,探讨其与肝硬化发生、发展的关系。方法将本课题组重组构建的含HCC-22-5cDNA序列表达质粒,经表达和纯化获得HCC-22-5的融合蛋白(MBP-HCC-22-5),免疫家兔制备出抗MBP-HCC-22-5多克隆抗体。经ELISA和Western blot鉴定抗体的免疫活性后,用免疫组化SABC法检测49例人肝硬化组织和30例正常人肝组织中HCC-22-5的表达。结果经ELISA和Western blot鉴定所制备的多克隆抗体可以与MBP-HCC-22-5特异性结合。HCC-22-5主要表达于肝细胞质中。HCC-22-5在肝硬化组织中的表达与性别有关,且与年龄呈正相关。结论成功制备了抗HCC-22-5的多克隆抗体;HCC-22-5的表达与肝硬化的发生、发展有关。
[ Objective] To Prepare anti-HCC-22-5 polyelonal antibody and study its relationship with progression of hepatic cirrhosis. [ Methods ] The MBP-HCC-22-5 fusion protein was expressed and purified from expressing plasmid that recombined in HCC-22-5 cDNA. The fusion protein was used to immunize rabbit to prepare the anti-MBP-HCC-22-5 pulyclonal antibody. The pulyclonal antibody was purified and identified by ELISA and Western blot. 49 cases of hepatic cirrhosis and 30 cases of normal liver tissues were used for detection. Immunohistochemical staining for HCC-22-5 was performed with SABC method on paraltln sections. The results associated with clinical data were analyzed. [ Results ] The polyclonal antibody was found to specifically bind to MBP-HCC-22-5 in ELISA and Western blot analysis. HCC-22-5 protein was found in the cytoplasm of hepatocytes. The expression of HCC-22-5 protein was related to gender and positively related to age in hepatic cirrhosis tissues. [ Conclusion] Anti-HCC-22-5 polyclonal antibody is prepared successfully. HCC-22-5 is associated with progression of hepatic cirrhosis.
出处
《山东医药》
CAS
北大核心
2007年第26期1-3,共3页
Shandong Medical Journal
基金
国家自然科学基金资助项目(No.39960022No.30460037)
高校博士点专项科研基金资助项目(No.20050598006)
