Surface plasmon resonance-based trace detection of small molecules by competitive and signal enhancement immunoreaction.
Surface plasmon resonance-based trace detection of small molecules by competitive and signal enhancement immunoreaction.
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DOI：
10.1016/j.aca.2007.03.074
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发表时间：
2007-05
影响因子：
6.2
通讯作者：
H. Aizawa;M. Tozuka;S. Kurosawa;K. Kobayashi;S. Reddy;M. Higuchi
中科院分区：
文献类型：
--
作者：
H. Aizawa;M. Tozuka;S. Kurosawa;K. Kobayashi;S. Reddy;M. Higuchi
研制了一种检测超低浓度低分子量化合物2，4-二硝基苯酚（DNP）的表面等离子体共振（SPR）免疫传感器。该传感器策略是基于DNP和DNP-蛋白质缀合物之间的竞争性免疫反应，即DNP-牛血清白蛋白缀合物（DNP-BSA）。利用化学偶联法将抗DNP单克隆抗体固定在金薄膜涂层SPR传感器芯片上。DNP-BSA在与抗-DNP抗体固定的SPR-免疫传感器芯片接触时引起传感器芯片的共振角增加。SPR传感器芯片上固定化抗体的最佳浓度为100μgmL−1。SPR-免疫传感器对游离DNP测定的响应时间约为20 min。15分钟。该方法可测定1 ppt ~ 1 ppb范围内的DNP。DNP的ppt水平的SPR信号通过随后用第二抗DNP抗体处理免疫结合的DNP-BSA增强三倍。
A surface plasmon resonance (SPR)-immunosensor for detection of the low molecular weight compound 2,4-dinitorophenol (DNP) at ultra-low concentration has been developed. The sensor strategy is based on a competitive immunoreaction between DNP and a DNP–protein conjugate, namely DNP–bovine serum albumin conjugate (DNP–BSA). Anti-DNP monoclonal antibody was immobilized on a gold thin-film coated SPR-sensor chip by means of a chemical coupling process. DNP–BSA, on contact with the anti-DNP antibody immobilized SPR-immunosensor chip causes an increase in the resonance angle of the sensor chip. The optimum concentration of immobilized antibody on the SPR-sensor chip is 100μgmL−1. The SPR-immunosensor response for free DNP determination using the competitive immunoreaction had a response time of ca. 15min. Using this method, DNP could be determined in the concentration range 1ppt to 1ppb. The SPR signal for ppt levels of DNP was enhanced by a factor of three by subsequently treating immuno-bound DNP–BSA with a secondary anti-DNP antibody.