MODULAR SURFACE PLASMON RESONANCE BIOSENSOR FOR IN-VITRO DETERMINATION OF PROTEIN

该项目将动电 (EK) 方法与表面等离子共振 (SPR) 光谱相结合，构建了一个灵活的模块化芯片实验室平台，用于确定目标生物标志物。小体积通道被蚀刻到介电基板上，其中使用 EK 方法将目标分析物与其他分析物进行浓缩和分离（即介电泳和电泳捕获） 生物流体中的化合物用于调节样品以进行 SPR 检测。例如，将采用介电泳和电泳捕获来局部浓缩微流体通道中的目标分析物，同时最小化干扰蛋白的局部浓度。金传感垫可用于直接无标记 SPR 分析，涂有生物受体以提高灵敏度 和选择性，或涂有生物受体进行分析，然后用针对目标生物标志物的另一种抗体进行信号放大。信号增强方法（例如抗体夹心测定）将包含在芯片实验室设计中，以实现更高程度的灵敏度。该传感器将应用于心肌梗塞生物标志物的检测。 拟议的平台具有以下对任何生物测定有益的属性： • 基于模块化“芯片”的传感平台可以分离、浓缩和定量面板检测所需的多种蛋白质和肽生物标志物。 • 动电法和夹心分析法的SPR 检测都适合分析蛋白质和肽的生理水平。 • 该系统非常适合小体积分析。 EP 和 DEP 在直径约为 10 至 100 微米的小通道中效果最佳。 SPR 对金传感垫 200 nm 范围内的折射率变化敏感。 • EK 可以根据电泳迁移率从复杂混合物中快速分离和浓缩分析物。可以实现高分辨率分离，并且可以通过介电泳或可寻址电泳捕获实现 1000 倍预浓缩。

This project combines electrokinetic (EK) methods with surface plasmon resonance (SPR) spectroscopy to construct a flexible, modular lab-on-a-chip platform for determination of target biomarkers. Small volume channels are etched into a dielectric substrate where EK methods for concentration and separation (i.e. dielectrophoresis and electrophoretic capture) of target analytes from the other compounds in the biofluids are employed to condition the sample for SPR detection. For example dielectrophoresis and electrophoretic capture will be employed to locally concentrate the target analytes in the microfluidic channel while minimizing the local concentration of interfering proteins. Gold sensing pads can be employed for direct label-free SPR analyses, coated with bioreceptors to increase sensitivity and selectivity, or coated with bioreceptors for the analysis followed by signal amplification with the another antibody to the target biomarker. Methods for signal enhancement such as antibody sandwich assays will be included with the lab-on-a-chip design to achieve a greater degree of sensitivity. This sensor will be applied to detection of biomarkers for myocardial infarction. The proposed platform offers the following attributes beneficial to any bioassay: • The modular, 'chip' based sensing platform can separate, concentrate, and quantify multiple protein and peptide biomarkers required for a panel assay. • Electrokinetic methods and SPR detection with sandwich assays are both appropriate for analyzing physiological levels of proteins and peptides. • The system is ideally suited for small volume analyses. EP and DEP work best in small channels approximately 10 to 100 microns in diameter. SPR is sensitive to refractive index changes within 200 nm of the gold sensing pads. • EK can rapidly separate and concentrate analytes from complex mixtures based on electrophoretic mobility. High resolution separations are possible and 1000-fold preconcentration can be achieved with dielectrophoresis or addressable electrophoretic capture.