铜完全抗原的合成·鉴定及单克隆抗体的制备  

The Synthesis and Identification of Copper Complete Antigen and the Preparation of Monoclonal Antibodies against Copper Ions

作　　者：刘文迪 周玉[1,2] 卢士英[1,2] 任洪林[1,2] 李岩松[1,2] 柳増善 宋芳 郝亚明 

机构地区：[1]吉林大学人兽共患病教育部重点实验室 [2]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《安徽农业科学》2011年第29期18072-18074,共3页Journal of Anhui Agricultural Sciences

基　　金：国家自然科学基金项目(60971011);吉林大学人才跟进计划项目(43050502J9)

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。[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.

关 键 词：CU2+ 抗原合成 鉴定 单克隆抗体 

分 类 号：X753[环境科学与工程—环境工程]