摘要
为研究杂环类双功能螯合剂在重金属铜人工抗原制备方面的效果,以2-S-(4-氨基苯)-1,4,7三氮杂环壬烷-1,4,7-三乙酸(P-NH_2-Bn-NOTA,简写为NOTA)为双功能螯合剂,采用戊二醛法将重金属铜离子分别与载体蛋白BSA、OVA偶联制备免疫原和包被原,并用合成的免疫原免疫制备免多克隆抗体,建立间接竞争ELISA(ic-ELISA)。结果表明,所建立的ic-ELISA线性回归方程为y=7.8632Ln(x)+9.6765(R2=0.9934),IC50和IC20值分别为168.70、3.72 ng·m L-1。交叉反应试验结果显示,除与锌交叉反应率达16%外,该抗体与其他重金属铁、铅和镉的交叉反应率均小于5.6%。本研究成功合成了重金属铜人工抗原并制备了兔多克隆抗体,为杂环类双功能螯合剂应用于重金属快速检测技术奠定了理论基础。
In order to study the effect of heterocyclic bifunctional chelators on the preparation of heavy metal copper artifical antigen. Using 2-S-（4-aminobenzene）-1,4,7 nitrogen heterocyclic nonane-1,4,7-triacetic acid（P-NH_2-Bn-NOTA or NOTA） as a bifunctional chelator,the heavy metal copper ions were conjugated with vector proteins BSA and OVA respectively to prepare immunogen and pactin,and the rabbit polyclonal antibodies were prepared by synthesis of immunogenic preparation,establishing indirect competition ELISA（ic-ELISA）. The results showed that the established linear regression equation of ic-ELISA was y = 7. 8632 Ln（x） ＋ 9. 6765（R^2= 0. 9934）,and the values of IC50 and IC20 were 168. 70 and 3. 72 ng·ml^-1,respectively. The cross-reaction test results showed that except the cross-reaction rate with zinc reached 16%,the cross-reaction rate between the antibody and other heavy metals iron,lead and cadmium was less than 5. 6%. In this study,heavy metal copper antigen was synthesized successfully and rabbit polyclonal antibody was prepared,which laid a thororetical foundation for the application to the rapid detection of heavy metal by heterocyclic bifunctional chelators.
作者
郭建军
桑丽雅
王振国
陈笑笑
金仁耀
GUO Jianjun;SANG Liya;WANG Zhenguo;CHEN Xiaoxiao;JIN Renyao(Key Laboratory of Aquatic Products Processing of Zhejiang Province/Institute of Seafood,Zhejiang Gongshang University,,Hangzhou,Zhejiang 310012;Nankai Biotechnology Limited company,Hangzhou,Zhejiang 310051)
出处
《核农学报》
CAS
CSCD
北大核心
2018年第10期2012-2018,共7页
Journal of Nuclear Agricultural Sciences
基金
国家自然科学青年基金(31501562)
浙江省科技厅公益性研究项目(2015C33093)
