Antibody-based sensors for heavy metal ions
Antibody-based sensors for heavy metal ions
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DOI：
10.1016/s0956-5663(01)00223-8
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发表时间：
2001-12-01
影响因子：
12.6
通讯作者：
Yu, HN
中科院分区：
文献类型：
--
作者：
Blake, DA;Jones, RM;Yu, HN
使用相同的试剂以两种不同的测定形式开发了Cd（II）、Co（II）、Pb（II）和U（VI）的竞争性免疫测定，竞争性微孔形式和KinExA（TM）3000的免疫传感器形式。四种不同的单克隆抗体特异性络合物的EDTA-Cd（II），DTPA-Co（II），2，9-二羧基-1，10-菲咯啉-U（VI），或环己基-DTPA-Pb（II）孵育适当的可溶性金属螯合物。在微孔测定形式中，金属螯合物络合物的固定化形式同时存在于测定混合物中。在KinExA形式中，使抗体与可溶性金属螯合物复合物预平衡，然后使孵育混合物快速通过含有固定化金属螯合物复合物的微柱。在所有四种测定中，KinExA形式产生具有10-1000倍更高灵敏度的测定。KinExA形式的灵敏度增强最有可能是由于单克隆抗体对可溶性金属螯合物与固定化金属螯合物的亲和力的差异。KinExA 3000仪器和Cd（II）特异性抗体被用来构建一个原型测定，可以正确地评估地下水样品中掺入镉的浓度。添加Cd（II）的平均分析回收率为114.25 +/-11.37%。该方法的精密度符合要求，批内和批间精密度的变异系数分别为0.81-7.77%和3.62-14.16%。(C)2001 Elsevier Science B. V.保留所有权利。
Competitive immunoassays for Cd(II), Co(II), Pb(II) and U(VI) were developed using identical reagents in two different assay formats, a competitive microwell format and an immunosensor format with the KinExA (TM) 3000. Four different monoclonal antibodies specific for complexes of EDTA-Cd(II), DTPA-Co(II), 2,9-dicarboxyl-1,10-phenanthroline-U(VI), or cyclohexyl-DTPA-Pb(II) were incubated with the appropriate soluble metal-chelate complex. In the microwell assay format, the immobilized version of the metal-chelate complex was present simultaneously in the assay mixture. In the KinExA format, the antibody was allowed to pre-equilibrate with the soluble metal-chelate complex, then the incubation mixture was rapidly passed through a microcolumn containing the immobilized metal-chelate complex. In all four assays, the KinExA format yielded an assay with 10-1000-fold greater sensitivity. The enhanced sensitivity of the KinExA format is most likely due to the differences in the affinity of the monoclonal antibodies for the soluble versus the immobilized metal-chelate complex. The KinExA 3000 instrument and the Cd(II)-specific antibody were used to construct a prototype assay that could correctly assess the concentration of cadmium spiked into a groundwater sample. Mean analytical recovery of added Cd(II) was 114.25 +/- 11.37%. The precision of the assay was satisfactory; coefficients of variation were 0.81-7.77% and 3.62-14.16% for within run and between run precision, respectively. (C) 2001 Elsevier Science B.V. All rights reserved.