| 抗人DcR3单克隆抗体的制备、鉴定及应用 |
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| 引用本文： | 刘瑞振,张长弓,何琼,苏金华,陈彩霞,陈福,庄国洪. 抗人DcR3单克隆抗体的制备、鉴定及应用[J]. 免疫学杂志, 2009, 25(1) |
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| 作者姓名： | 刘瑞振 张长弓 何琼 苏金华 陈彩霞 陈福 庄国洪 |
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| 作者单位： | 刘瑞振,张长弓,苏金华,陈彩霞,陈福,庄国洪(厦门大学医学院抗癌研究中心,厦门,361005);何琼(厦门市波生生物技术有限公司) |
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| 基金项目： | 厦门大学科研启动资金资助项目，厦门市科技计划项目 |
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| 摘 要： | 目的 制备抗DcR3单克隆抗体(mAb),鉴定其生物学特性,并应用于ELISA、Western blot、How cytometry(FCM)检测.方法 以纯化的可溶性DcR3(sDcR3)免疫Balb/c小鼠,采用杂交瘤技术制备抗DcR3 mAb.用Ig亚类EHSA试剂盒鉴定抗DcR3mAb的亚类.用ELISA方法测定抗DcR3 mAb与sDcR3结合的特性,SDS-PAGE鉴定抗DcR3 mAb与SW480细胞上清中DcR3结合的特性,以鉴定mAb的特性.用间接ELISA法检测腹水mAb的效价、亲和常数并进行表位分析.Western blot检测mAb的特异性及应用,并用所获抗DcR3单克隆抗体(mAb)通过流式细胞仪检测肿瘤细胞表面DcR3的表达水平.结果 获得4株可分泌DcR3 mAb的杂交瘤细胞系ZZ-393、ZZ-394、ZZ-151和ZZ-268.其中DcR3 mAb ZZ-268(下文简称为ZZ-268)的Ig亚类为IgGl(k型);腹水效价为1×10-5;亲和常数为1.28×109水平;ZZ-268和ZZ-151可识别与其他2种抗体小同的抗原表位;Westem blot证实,获得的ZZ-268可特异性地识别DcR3;通过流式细胞术可敏感地检测到不同肿瘤细胞表面DcR3的表达水平.结论 获得4株抗DcR3的mAb,其中ZZ-268效价高、特异性强,将此抗体用于膜DcR3与sDcR3的检测分析.
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| 关 键 词： | 单克隆抗体 制备 |
Preparation,characterization,and application of monoclonal antibodies against human decoy receptor 3 |
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| LIU Ruizhen,ZHANG Changgong,HE Qiong,CHEN Caixia,CHEN Fu,ZHUANG Guohong Anti-Cancer Research Center,Medical College,Xiamen University,Xiamen ,China. Preparation,characterization,and application of monoclonal antibodies against human decoy receptor 3[J]. Immunological Journal, 2009, 25(1) |
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| Authors: | LIU Ruizhen ZHANG Changgong HE Qiong CHEN Caixia CHEN Fu ZHUANG Guohong Anti-Cancer Research Center Medical College Xiamen University Xiamen China |
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| Affiliation: | LIU Ruizhen,ZHANG Changgong,HE Qiong,CHEN Caixia,CHEN Fu,ZHUANG Guohong Anti-Cancer Research Center,Medical College,Xiamen University,Xiamen 361005,China |
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| Abstract: | Objective To prepare monoclonal antibodies mAb against DcR3,characterize their biological properties and apply them to ELISA,Flow cytometry,and Western blot detection.Methods Balb/c mice were immunized with soluble DcR3(sDcR3) for preparing mAb by using hybridoma technique.Ig subclass and specificity of mAbs were analyzed by ELISA and SDS-PAGE,respectively.The titres,relative affinity,and epitopes of mAbs in ascitic fluid were determined by indirect ELISA.The specificity and application of mAb were detected... |
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| Keywords: | DcR3 |
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