摘要
目的:探究纳米SiO_(2)富集分离水样中Cd^(2+)的条件,制备抗Cd^(2+)单克隆抗体,建立水样中痕量Cd^(2+)异源性间接竞争ELISA(heterologous indirect competitive ELISA,hic ELISA)快速检测方法。方法:以纳米SiO_(2)富集、EDTA-2Na竞争洗脱Cd^(2+),ICP法测定处理过程中的Cd^(2+)含量。人工合成Cd-ITCBE-BSA免疫Balb/C小鼠,制备Cd^(2+)单克隆抗体。人工合成异源性包被抗原,建立镉离子hic ELISA检测方法。结果:纳米SiO_(2)对Cd^(2+)的吸附容量为13.3 mg/g,富集倍数可达20倍,EDTA-2Na可有效洗脱被富集的Cd^(2+)。以Cd-ITCBE-BSA为免疫原,小鼠血清效价达到1:1.28×10^(4);腹水效价为1:2.0×10^(5),亲和力Ka为8.1×10^(8) L/moL。以Hg-ITCBE-OVA包板建立的hic ELISA检测限为2.9μg/L,与Hg^(2+)、Cu^(2+)、Zn^(2+)、Pb^(2+)、Cr^(3+)、Mo^(6+)、Fe^(3+)、Co^(2+)无明显交叉反应。在湖水、自来水、超纯水样品中镉的加标回收率为92.47%~102.86%。结论:建立的纳米SiO_(2)富集重金属镉-hicELISA检测方法灵敏、特异、准确,能用于水样中镉离子的检测。
Objective:Explore the conditions of preconcentration and separation of Cd^(2+)in water samples by nano-SiO_(2),prepare monoclonal antibody against Cd^(2+) and establish a rapid detection method of trace Cd^(2+) in water samples by heterologous indirect competitive ELISA(hicELISA).Methods:Cd^(2+)was preconcentrated by nano-SiO_(2)and eluted by EDTA-2 Na.The content of Cd^(2+) was determined by ICP.Balb/C mice were immunized with Cd-ITCBE-BSA to prepare Cd^(2+) monoclonal antibody.The heterologous coating antigen was synthesized to establish the detection method of cadmium ion hicELISA.Results:The adsorption capacity of Nano-SiO_(2)for Cd^(2+) was 13.3 mg/g,and the enrichment multiple was 20 times.EDTA could be used as the effective eluent for the pretreatment.Using Cd-ITCBE-BSA as immunogen,the titer of serum was 1:1.28×10^(4),The titer of mAb in ascites was 1:2.0×10^(5)and the affinity constantwas 8.1×10^(8)L/mol.The detection limit of hicELISA was 2.9μg/L.There was no significant cross reaction with Hg^(2+),Cu^(2+),Zn^(2+),Pb^(2+),Cr^(3+),Mo^(6+),Fe^(3+),CO_(2)+.The recoveries of cadmium in lake water,tap water and ultrapure water were 92.47%~102.86%.Conclusion:The hicELISA method was sensitive,specific and accurate,and could be used for the detection of cadmium in water samples.
作者
周叶
杨树宇
高祎琳
陈俊杰
刘长忠
王自良
姜金庆
ZHOU Ye;YANG Shuyu;GAO Yilin;CHEN Junjie;LIU Changzhong;WANG Ziliang;JIANG Jinqing(College of Animal Science and Veterinary Medicine,Henan Institute of Science and Technology,Xinxiang 453003,China)
出处
《食品工业科技》
CAS
北大核心
2021年第14期278-283,共6页
Science and Technology of Food Industry
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)。
