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抗人可溶性间皮素相关蛋白单克隆抗体的制备及特性鉴定
引用本文:陈书成,刘雅坤,吴小华,单保恩,陈兴,孙东霞,王莉. 抗人可溶性间皮素相关蛋白单克隆抗体的制备及特性鉴定[J]. 细胞与分子免疫学杂志, 2007, 23(3): 246-248
作者姓名:陈书成  刘雅坤  吴小华  单保恩  陈兴  孙东霞  王莉
作者单位:1. 河北医科大学第四医院妇产科,河北,石家庄,050011
2. 河北医科大学第四医院科研中心,河北,石家庄,050011
3. 白求恩国际和平医院,河北,石家庄,050000
基金项目:国家自然科学基金;河北省自然科学基金
摘    要:目的:制备鼠抗人可溶性间皮素相关蛋白(SMR)的单克隆抗体(mAb),鉴定其生物学特性。方法:应用计算机通过综合预测对间皮素(MSLN)进行B-细胞表位预测。合成相应肽段并免疫BALB/c小鼠,采用杂交瘤技术制备mAb,利用免疫细胞化学和Western blot分析对所制备的mAb进行鉴定。结果:综合预测方法分析间皮素的抗原表位可能于153-162、282-292及471-481氨基酸残基或其附近,选择性合成了可能性最高的位于471-481氨基酸残基的可溶性间皮素相关蛋白表位,制备了mAb(2H10),经免疫细胞化学和Western blot分析该抗体具有较高的特异性。结论:成功地制备了鼠抗人可溶性间皮素相关蛋白的mAb(2H10),该抗体具有较高的特异性,为进一步利用ELISA对可溶性间皮素相关蛋白进行检测奠定了基础。

关 键 词:间皮素  可溶性间皮素相关蛋白  B细胞表位  单克隆抗体
文章编号:1007-8738(2007)03-0246-03
修稿时间:2006-12-05

Preparation and characterization of the monoclonal antibody against human soluble mesothelin-related proteins
CHEN Shu-cheng,LIU Ya-kun,WU Xiao-hua,SHAN Bao-en,CHEN Xing,SUN Dong-xia,WANG Li. Preparation and characterization of the monoclonal antibody against human soluble mesothelin-related proteins[J]. Chinese journal of cellular and molecular immunology, 2007, 23(3): 246-248
Authors:CHEN Shu-cheng  LIU Ya-kun  WU Xiao-hua  SHAN Bao-en  CHEN Xing  SUN Dong-xia  WANG Li
Affiliation:1.Department of Obstetrics and Gynaecology, the Fourth Hosipital of Hebei Medical University, 2 Research Center of the Fourth Hosipital of Hebei Medical University, Shijiazhuang 050011; 3.Bethune International Peace Hosipital, Shijiazhuang 050000, China
Abstract:AIM:To prepare the monoclonal antibody(mAb) against human soluble mesothelin-related proteins(SMR) and identify its properties.METHODS: The B-cell epitopes of mesothelin(MSLN) were predicted by multi-parameter prediction method.Then BALB/c mice were immunized with compound MSLN polypeptide prepare the mAb by hybridoma technique.The specificity of antibody was evaluated by immuocytochemistry and Western blot respectively.RESULTS: Using the multi-parameter prediction of B-cell epitopes,the 471-481 epitope domain was selected and synthesized.Then the mice were immunized and mAb against human SMR was prepared and designated 2H10.The specificity of mAb 2H10 was evaluated to be high by using immuocytochemistry and Western blot.CONCLUSION: The successful preparation of the mAb against SMR will provide efficient reagent for further detection of SMR by ELISA.
Keywords:mesothelin  soluble mesothelin-related proteins  B-cell epitopes  monoclonal antibody
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