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Position: Home > Articles > Preparation of artificial immunogen of mercury ion and the identification of its immunological characteristics Heilongjiang Animal Science and Veterinary Medicine 2017 (7) 180-183+187

Hg~(2+)人工免疫原的制备及免疫特性鉴定

作者：

秦保亮;姜金庆;孙勇;付世杰;王亚楠

单位：

甘肃省治沙研究所;河南科技学院动物科学学院;河南科技学院;河南科技大学

关键词：

Hg2+;半抗原;人工免疫原;制备;酶联免疫吸附法(ELISA);免疫特性鉴定

摘要：

为了建立重金属汞离子(Hg~(2+))免疫学检测方法,试验采用异硫氰酯法将双功能螯合剂ITCBE与Hg~(2+)螯合形成半抗原,然后将其与牛血清白蛋白(BSA)和卵清蛋白(0VA)耦联合成人工免疫原Hg~(2+)-ITCBE-BSA和包被原Hg~(2+)-ITCBE-OVA,用紫外扫描分析(UV)、电感耦合等离子体质谱法(ICP-MS)和凝胶电泳(SDS-PAGE)进行鉴定;用Hg~(2+)-ITCBE-BSA免疫Balb/c小鼠,间接ELISA法测定多克隆抗体(p Ab)效价,阻断ELISA法鉴定其敏感性,交叉反应试验鉴定其特异性。结果表明:免疫原Hg~(2+)-ITCBE-BSA和包被原Hg~(2+)-ITCBE-OVA制备成功,Hg~(2+)-ITCBEBSA中BSA和Hg~(2+)的含量分别为9.09 g/L和45.88 mg/L;间接ELISA法检测效价达到1∶1.024×105,阻断ELISA法检测Hg-EDTA的IC50为45.49 g/L。说明Hg~(2+)人工免疫原制备成功。

译名：

Preparation of artificial immunogen of mercury ion and the identification of its immunological characteristics

作者：

QIN Baoliang;JIANG Jinqing;SUN Yong;FU Shijie;WANG Yanan;FAN Guoying;WANG Ziliang;College of Animal Science,Henan Institute of Science and Technology;Animal Disease Prevention and Control Center of Xinxiang City;

关键词：

Hg2+;;hapten;;artificial immunogen;;preparation;;enzyme-linked immunosorbent assay(ELISA);;identification of immunological characteristics

摘要：

To establish a method of immunological detection of heavy metal mercury ion,a haptene was formed by the chelating agent ITCBE and mercury ion using the method of isothiocyanate.The haptene was coupled and synthesized with bovine serum albumin(BSA) and ovalbumin(OVA) into artificial immunogen Hg~(2+)-ITCBE-BSA and coating antigen Hg~(2+)-ITCBE-OVA,while the ultraviolet(UV) scanning analysis,inductively coupled plasma mass spectrometry(ICP-MS) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) were used for the identification.Balb/c mice were immunized with Hg~(2+)-ITCBE-BSA,the titer of polyclonal antibody(p Ab) was detected by indirect ELISA,and the sensitivity of p Ab was identified by blocking ELISA and the specificity of p Ab was identified by cross-reaction test.The results showed that the immunogen Hg~(2+)-ITCBE-BSA and the coating antigen Hg~(2+)-ITCBE-OVA were successfully prepared.The contents of BSA and Hg~(2+)in Hg~(2+)-ITCBE-BSA were 9.09 g/L and 45.88 mg/L.The titer of Hg~(2+)-EDTA p Ab of Balb/c mice reached 1 ∶ 1.024 × 105,which was detected by indirect ELISA,and the 50% inhibitive concentration(IC50) of Hg-EDTA was45.49 g/L,which was detected by blocking ELISA.The results indicate that the artificial immunogen of heavy mercury ion is successfully prepared.

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Hg~(2+)人工免疫原的制备及免疫特性鉴定

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