【作者】 袁晓科； 杨志强； 李建喜； 王学智； 陈化琦； 张艳； 孟嘉仁； 高杰；

【Author】 YUAN Xiao-ke YANG Zhi-qiang LI Jian-xi WANG Xue-zhi CHEN Hua-qi ZHANG Yan MENG Jia-ren GAO Jie(Lanzhou Institute of Animal Sciences and Veterinary Pharmaceutics of CAAS,Gansu Lanzhou 730050)

【机构】 中国农业科学院兰州畜牧与兽药研究所；

【摘要】 为了建立重金属镉离子ELISA快速检测技术、制备单克隆抗体,本研究选用螯合剂Isothiocyanobenzyl-EDTA (IEDTA)分别桥接载体蛋白KLH和BSA,络合重金属镉离子。制备免疫抗原和检测抗原,免疫Balb/C小鼠并制备抗血清;利用紫外分光光度法、三硝基苯磺酸法(TNBS)和测定抗体效价。结果表明,抗原紫外吸收峰与载体蛋白紫外吸收峰有明显差异,两种抗原游离氨基酸含量在33%-40%之间,抗原蛋白含量均高于70%,血清效价为1:81920以上。结果提示,免疫抗原特异性较好,可用于ELISA检测技术研究和单克隆抗体制备。更多还原

【Abstract】 In order to study ELISA method(a kind of rapid detection)and to prepare monoclonal antibody on heavy metal ion cadmium.The immunizing antigen and detecting antigen were synthesized with isothiocyanobenzyl-EDTA(IEDTA),carrier protein KLH or BSA and cadmium nitrate.Balb/C mice were immunized with immunizing antigen to obtain the antiserum.Two antigens were identified by three methods including ultraviolet spectrophotometry,trinitro-benzene-sulfonic acid(TNBS)and the antiserum titer of mice.The different ultraviolet absorbance was observed between synthesized antigen and carrier protein,the content of free amino acid in antigen was from 33% to 40%,the titer of antiserum was above 1:81920.These results indicated that the immunizing antigen was obvious specificity and it is an ideal candidate of immunizing antigen for preparation of monoclonal antibody and for researching of ELISA detecting method更多还原