【摘要】 制备并鉴定针对重金属镉离子的小鼠多克隆抗体.选取异硫氰酸苄基乙二胺四乙酸（ITCBE）做金属螯合剂,偶联Cd²⁺及载体蛋白BSA、OVA,制备人工抗原并鉴定,配合佐剂乳化后以高、中、低剂量分别免疫Balb/C小鼠,以获得较好的抗重金属镉离子小鼠多克隆抗体血清.采用酶联免疫吸附法（ELISA）测定所获抗体的效价、敏感度、特异性.结果显示:小鼠抗血清效价均在10^-3～10^-4,中剂量抗原免疫组小鼠抗体血清更加稳定、特异性强,利用该组多抗建立间接竞争ELISA（icELISA）标准曲线,M-3号小鼠敏感性最强,回归方程为y=0.1123x-0.1638,IC₅₀质量浓度为368.70μg/L,检测限质量浓度为25.53μg/L,该抗体除与汞离子螯合物的交叉反应率为72.58%之外,与Cu²⁺、Zn²⁺、Pb²⁺、Cr³⁺、Mo⁶⁺、Fe³⁺、Co²⁺均无交叉反应.获得了能用于镉离子免疫检测的高效价Cd²⁺多抗,为检测产品的开发打下基础.更多还原

【Abstract】 To prepare and identify mouse polyclonal antibodies（PcAbs） against cadmium,using bifunctional chelating agent ITCBE to connect Cd²⁺with carrier proteins BSA and OVA for the preparation of artificial antigen.Balb/C mice were immunized with high,medium and low doses of antigen emulsified by adjuvant to obtain better polyclonal antibody serum.The titer,sensitivity and specificity of the antibody were determined by enzyme linked immunosorbent assay（ELISA） methods.The titers of mouse antiserum ranged from 10^-3 to 10^-4.The antibody serum of mice immunized with medium dose antigen was more stable and specific.The standard curve of indirect competitive ELISA（icELISA） was established by using the antibody.The IC₅₀ value was 368.70 μg/L,and the detection limit was25.53 μg/L.The antibody had no cross reaction with Cu²⁺,Zn²⁺,Pb²⁺,Cr³⁺,Mo⁶⁺,Fe³⁺,Co²⁺except the cross reaction rate with mercury ion chelate was 72.58%.Cd²⁺polyclonal antibodies with high titer were obtained,which laid a foundation for the development of detection products.更多还原