Multichannel surface plasmon resonance imaging and analysis of micropatterned self-assembled monolayers and protein affinity interactions
Multichannel surface plasmon resonance imaging and analysis of micropatterned self-assembled monolayers and protein affinity interactions
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DOI：
10.1021/la0486382
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发表时间：
2005-01-04
期刊：
影响因子：
3.9
通讯作者：
Yoon, HC
中科院分区：
文献类型：
--
作者：
Pyo, HB;Shin, YB;Yoon, HC
利用二维表面等离子体共振（2D-SPR）成像系统，观察了11-巯基十烷酸和11-巯基十一烷醇自组装单层膜与生物特异性干扰素- γ (ifn - γ)/抗lfn - γ抗体免疫亲和相互作用的多通道图像。利用自制的2D-SPR成像系统，采用白光光源结合窄带通滤光片，对清晰图像进行了分辨，最大限度地减少了成像图像上的衍射图样。利用巯基键的紫外光解取代传统的光刻技术，实现了自组装单层的微图图化。利用椭圆分析法对图像进行线轮廓校正，使我们能够在亚纳米尺度内区分单层厚度的变化。对于蛋白在表面的相互作用，分析了ifn - γ抗原与表面固定化抗体的生物特异性亲和识别反应。在三明治型免疫分析中，通过基于酶催化沉淀反应的信号放大策略，可以检测到低至1 ng/mL的生物特异性抗原结合。
Multichannel images of 11-mercaptoundecanoic acid and 11-mercapto-1-undecanol self-assembled monolayers together with a biospecific interferon-gamma (IFN-gamma)/anti-lFN-gamma antibody immunoaffinity interaction were observed by the two-dimensional surface plasmon resonance (2D-SPR) imaging system. With the fabricated 2D-SPR imaging system, adopting a white light source in combination with a narrow band-pass filter, sharp images were resolved, minimizing the diffraction patterns on the resulting images. Micropatterning of self-assembled monolayers was acheived by exploiting the UV photolysis of thiol bonding, instead of conventional photolithography. The line profile calibration of the image contrast with ellipsometric analysis enabled us to discriminate the change in monolayer thickness within a sub-nanometer scale. For the protein interactions on the surface, the biospecific affinity recognition reaction of IFN-gamma antigen with surface-immobilized antibody was analyzed. Through the signal amplification strategy based on the enzyme-catalyzed precipitation reaction in a sandwich-type immunoassay, biospecific antigen binding was found detectable down to a concentration of 1 ng/mL.