Plasma-enhanced antibody immobilization for the development of a capillary-based carcinoembryonic antigen immunosensor using laser-induced fluorescence spectroscopy.
Plasma-enhanced antibody immobilization for the development of a capillary-based carcinoembryonic antigen immunosensor using laser-induced fluorescence spectroscopy.
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DOI：
10.1021/ac400226n
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发表时间：
2013-04
影响因子：
7.4
通讯作者：
Qiaoling Yu;Xuefang Zhan;Kunping Liu;Hao Lv;Y. Duan
中科院分区：
文献类型：
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作者：
Qiaoling Yu;Xuefang Zhan;Kunping Liu;Hao Lv;Y. Duan
在这项研究中，使用微波诱导的H2O/Ar等离子体预处理的抗体固定化首次实现。使用等离子体通过增加硅羟基和悬挂键的密度来活化基于毛细管的免疫传感器的表面，以确保更好的硅烷化。在硅烷化表面与戊二醛和抗体反应后，共价固定捕获抗体。将Cy 3标记的检测抗体与免疫传感器捕获的抗原组合使用以完成免疫印迹型免疫测定，并使用激光诱导荧光系统测量信号。微波诱导的H2O/Ar等离子体预处理癌胚抗原（CEA）免疫传感器提高了抗体的固定化，线性检测范围有明显提高，与商业酶联免疫吸附测定（ELISA）相比，1个数量级。这种新的固定方法显着提高了检测限（0.5 pmol/L CEA）和灵敏度。方法的相关系数为0.9978，相对标准偏差<7%，回收率为99.7- 107.1%。该免疫传感器成功应用于实际唾液标本中CEA的测定，具有较高的灵敏度、可接受的精密度和合理的准确度。这种基于微波诱导H2O/Ar等离子体的增强型CEA免疫传感器被证明是CEA诊断的敏感工具。
In this study, antibody immobilization using a microwave-induced H2O/Ar plasma pretreatment was achieved for the first time. Plasma was used to activate the surface of a capillary-based immunosensor by increasing the density of silicon hydroxyls and dangling bonds to ensure better silanization. The capture antibodies were covalently immobilized after the silanized surface reacted with glutaraldehyde and antibodies. A Cy3-labeled detection antibody was used in combination with the antigen captured by the immunosensor to complete the sandwich-type immunoassay, and the signals were measured using a laser-induced fluorescence system. Microwave-induced H2O/Ar plasma pretreatment of the carcinoembryonic antigen (CEA) immunosensor improved the antibody immobilization, and there was an obvious improvement in the linear detection range, i.e., 1 order of magnitude compared with a commercial enzyme-linked immunosorbent assay (ELISA). This novel immobilization method dramatically improved the detection limit (0.5 pmol/L CEA) and sensitivity. Assay validation studies indicated that the correlation coefficient reached 0.9978, and the relative standard deviations were <7% for all samples, with recoveries of 99.7-107.1%. Furthermore, the immunosensor was applied successfully to CEA determination in actual saliva specimens with high sensitivity, acceptable precision, and reasonable accuracy. This enhanced CEA immunosensor based on microwave-induced H2O/Ar plasma was demonstrated to be a sensitive tool for CEA diagnostics.