Preparation of a novel monoclonal antibody specific for WIG-1 and detection of its expression pattern in human esophageal carcinoma.
Preparation of a novel monoclonal antibody specific for WIG-1 and detection of its expression pattern in human esophageal carcinoma.
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DOI：
10.1089/hyb.2010.0043
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发表时间：
2010-11
期刊：
影响因子：
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通讯作者：
Ying-Bo Zou;Wei Guo;Yao-guang Jiang;Ru-wen Wang;Yun-ping Zhao;Zheng Ma
中科院分区：
文献类型：
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作者：
Ying-Bo Zou;Wei Guo;Yao-guang Jiang;Ru-wen Wang;Yun-ping Zhao;Zheng Ma
我们已经有效地产生了第一个单克隆抗体（单克隆抗体）对人类WIG-1（野生型p53诱导基因1）蛋白，一个骨化相关蛋白组成的三个锌指结构域。用蛋白A亲和层析法从小鼠腹水中纯化单克隆抗体。用间接酶联免疫吸附试验（ELISA）筛选杂交瘤。用ELISA和Western免疫印迹法对一株特异性单克隆抗体GW-3E 4（IgG 1）进行了鉴定，该单克隆抗体能有效地检测人食管鳞癌（ESCC）组织和EC 109细胞的核蛋白。因此，它结合天然WIG-1蛋白，并应在WIG-1蛋白的功能和表达的研究是有用的。用该单克隆抗体对20例食管鳞癌患者进行Western免疫印迹分析，发现WIG-1蛋白在食管鳞癌组织中的表达明显高于切缘组织。结果表明，WIG-1可能是食管癌发生过程中一种新的调节因子，WIG-1单克隆抗体的制备为进一步研究WIG-1相关生理反应的机制奠定了基础。
We have efficiently generated the first monoclonal antibody (MAb) against the human WIG-1 (wild-type p53-induced gene 1) protein, an apoptosis-related protein consisting of three zinc finger domains. Protein A affinity chromatography was performed to purify MAb from mice production ascites. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA). One MAb designated GW-3E4 (IgG1), effective in detecting the nuclear protein in human esophageal squamous cell carcinoma (ESCC) tissues and EC109 cell line, was characterized by ELISA and Western immunoblotting. Thus, it binds to native WIG-1 protein and should be useful in studies of WIG-1 protein function and expression. By Western immunoblotting analysis of 20 patients with ESCC using the MAb, we found that the expression of WIG-1 protein in tumor tissue was significantly higher than in incision margin. The results showed that WIG-1 might be a novel modifier in esophageal carcinogenesis, and the WIG-1 MAb should be useful in further study of the mechanism in WIG-1-related physiological reactions.