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抗软骨藻酸单克隆抗体杂交瘤细胞株的构建
引用本文:高利利,刘元嫄,程金平,王茜,王文华.抗软骨藻酸单克隆抗体杂交瘤细胞株的构建[J].海洋渔业,2010,32(3).
作者姓名:高利利  刘元嫄  程金平  王茜  王文华
作者单位:上海交通大学环境科学与工程学院,上海,200240
基金项目:上海市科委科技攻关项目 
摘    要:为了获得稳定、高效分泌抗软骨藻酸(DA)单克隆抗体的杂交瘤细胞株,采用活泼酯法制备软骨藻酸免疫抗原(DA-KLH)和包被抗原(DA-BSA),以DA-KLH免疫BALB/c小鼠,用细胞融合技术筛选抗软骨藻酸单克隆抗体杂交瘤细胞株,体内诱生腹水法制备大量单抗,捕获EL ISA法测定小鼠免疫球蛋白亚型,间接ELISA法测定腹水效价;用G蛋白亲和层析法来纯化腹水。结果显示,成功构建2株能稳定分泌DA单克隆抗体(McAb)的杂交瘤细胞株2B1和4C2,抗体亚类分别为IgG1和IgG2a,间接EL ISA检测小鼠腹水的抗体效价达1∶64 000,腹水纯化后蛋白浓度为1.27和0.675 mg/mL。研究结果表明,成功制备的抗DA单克隆抗体杂交瘤细胞株稳定、高效,为利用该单抗研制快速检测软骨藻酸的胶体金免疫层析试纸条打下基础。

关 键 词:软骨藻酸  单克隆抗体  杂交瘤细胞株

Preparation of hybridoma cell strains secreting monoclonal antibodies against domic acid
GAO Li-li,LIU Yuan-yuan,CHENG Jin-ping,WANG Qian,WANG Wen-hua.Preparation of hybridoma cell strains secreting monoclonal antibodies against domic acid[J].Marine Fisheries,2010,32(3).
Authors:GAO Li-li  LIU Yuan-yuan  CHENG Jin-ping  WANG Qian  WANG Wen-hua
Institution:GAO Li-li,LIU Yuan-yuan,CHENG Jin-ping,WANG Qian,WANG Wen-hua (Department of Environmental Science and Engineering,Shanghai Jiao Tong University,Shanghai 200240,China)
Abstract:To obtain stable and efficient hybridoma cell strains secreting monoclonal antibody(McAb) against domic acid(DA),completed antigen(DA-KLH) and coating antigen(DA-BSA) were synthesized by active ester method.BALB/c mice were immunized with DA-KLH and hybridoma cell strains secreting McAb against DA were screened by cell fusion,and the immunological titers of DA-McAb were determined.Capture ELISA method was used to determine isotypes of the mouse monoclonal antibody and the indirect ELISA method was performed...
Keywords:Domic acid  Monoclonal antibody  Hybridoma cell strains  
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