| 铜完全抗原的合成·鉴定及单克隆抗体的制备 |
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| 引用本文： | 刘文迪,周玉,卢士英,任洪林,李岩松,柳增善,宋芳,郝亚明.铜完全抗原的合成·鉴定及单克隆抗体的制备[J].安徽农业科学,2011,39(29):18072-18074. |
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| 作者姓名： | 刘文迪 周玉 卢士英 任洪林 李岩松 柳增善 宋芳 郝亚明 |
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| 作者单位： | 吉林大学人兽共患病教育部重点实验室,吉林大学畜牧兽医学院,吉林长春130062 |
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| 基金项目： | 国家自然科学基金项目(60971011);吉林大学人才跟进计划项目(43050502J9) |
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| 摘 要： | 目的]获得抗Cu2＋的单克隆抗体。方法]以异硫氰酸苄基乙二胺四乙酸（ITCBE）为双功能螯合剂,使Cu2＋分别与载体蛋白牛血清白蛋白（BSA）、卵清蛋白（OVA）偶联,获得免疫原（Cu-ITCBE-BSA）和检测原（Cu-ITCBE-OVA）。用非变性聚丙烯凝胶电泳鉴定偶联结果。免疫Balb/C小鼠,通过细胞融合,获得能稳定分泌抗Cu2＋的杂交瘤细胞株,并通过亲和层析法纯化腹水获得抗Cu2＋的单克隆抗体。结果]获得了1株能稳定分泌抗Cu2＋的单克隆抗体的细胞株（4A6）,所分泌的抗体亚类为IgG1,细胞培养上清效价可达1∶1.0×103,腹水效价可达1∶6.4×105。以该细胞株接种Balb/C小鼠腹腔,获得抗Cu2＋的腹水型单抗;该腹水经过亲和层析法纯化后,纯度可达95%。结论]获得了抗Cu2＋的单克隆抗体,为环境水样中Cu2＋的免疫学检测奠定基础。
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| 关 键 词： | Cu2＋ 抗原合成 鉴定 单克隆抗体 |
The Synthesis and Identification of Copper Complete Antigen and the Preparation of Monoclonal Antibodies against Copper Ions |
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| Institution: | LIU Wen-di et al(Key Laboratory of Zoonosis Research,Ministry of Education Institute of Zoonosis,College of Animal Science and Veterinary Medicine,Jilin University,Changchun,Jilin 130062) |
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| Abstract: | Objective] The aim was to gain the monoclonal antibody against copper ions.Method] The carrier protein BSA,OVA were separately chelated with bifunctional chelating agent Isothiocyanobenzyl-EDTA(ITCBE) and linked with ion copper to gain the completed Immunogen(Cu-ITCBE-BSA) and the detection Immunogen(Cu-ITCBE-OVA).Non-denaturing polyacrylamide gel electrophoresis was used for the identification of the completed antigen.By using cell fusion and limiting dilution,a stable hybridoma strains secreting anti Cu2+ was obtained.Result] A stable hybridoma strains(4A6) secreting anti Cu2+ monoclonal antibody was obtained.The sub-class of antibody was IgG1.The titer of cell culture supernatant and ascites were 1∶1024 and 1∶6.4×105 respectively.The ascites was purified by affinity chromatography,the purity was 95%.Conclusion] A monoclonal antibody against copper ions has been obtained,which provided foundations for the detection of copper ions in environmental water samples. |
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| Keywords: | Copper Antigen synthesis Identification Monoclonal antibody |
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