抗重金属镉离子鼠源性多克隆抗体的制备与鉴定  

Preparation and identification of mouse polyclonal antibody against heavy metal cadmium

作　　者：周叶 杨树宇 陈俊杰[1] 刘长忠[1] 王自良[1] 姜金庆[1] ZHOU Ye;YANG Shuyu;CHEN Junjie;LIU Changzhong;WANG Ziliang;JIANG Jinqing(College of Animal Science and Veterinary Medicine,Henan Institute of Science and Technology,Xinxiang 453003,China)

机构地区：[1]河南科技学院动物科技学院,河南新乡453003

出　　处：《河南科技学院学报（自然科学版）》2021年第1期24-29,共6页Journal of Henan Institute of Science and Technology(Natural Science Edition)

基　　金：中原千人计划-中原科技创新领军人才项目(204200510010);河南省高校科技创新团队(20IRTSTHN025);河南省重点科技攻关项目(202102110090)。

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Cd^(2+)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Cu^(2+)、Zn^(2+)、Pb^(2+)、Cr^(3+)、Mo^(6+)、Fe^(3+)、Co^(2+)均无交叉反应.获得了能用于镉离子免疫检测的高效价Cd^(2+)多抗,为检测产品的开发打下基础.To prepare and identify mouse polyclonal antibodies(PcAbs)against cadmium,using bifunctional chelating agent ITCBE to connect Cd^(2+)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 368.70μg/L,and the detection limit was 25.53μg/L.The antibody had no cross reaction with Cu^(2+),Zn^(2+),Pb^(2+),Cr^(3+),Mo^(6+),Fe^(3+),Co^(2+)except the cross reaction rate with mercury ion chelate was 72.58%.Cd^(2+)polyclonal antibodies with high titer were obtained,which laid a foundation for the development of detection products.

关 键 词：重金属 镉离子 多克隆抗体 鉴定 

分 类 号：S859.84[农业科学—临床兽医学]