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

该项目将电动(EK)方法与表面等离子体共振(SPR)光谱相结合，构建了一个灵活、模块化的芯片实验室平台，用于确定目标生物标记物。小体积通道被蚀刻到介质衬底中，其中EK方法用于目标分析物的浓缩和分离(即介电和电泳法捕获) 生物体液中的化合物被用来调节样品以进行SPR检测。例如，将使用介电泳法和电泳法捕获微流控通道中的目标分析物，同时将干扰蛋白质的局部浓度降至最低。金传感垫可用于直接的无标记SPR分析，涂有生物感受器以提高灵敏度 和选择性，或包被生物受体进行分析，然后用另一种抗体对靶生物标记物进行信号放大。芯片实验室设计将包括抗体夹心分析等信号增强方法，以实现更高程度的灵敏度。该传感器将应用于心肌梗死生物标志物的检测。 建议的平台提供了对任何生物检测有益的以下属性： ·基于芯片的模块化传感平台可以分离、浓缩和量化小组分析所需的多个蛋白质和多肽生物标记物。 ·电动方法和夹心法SPR检测都适用于分析蛋白质和多肽的生理水平。 ·该系统非常适合小量分析。EP和DEP在直径约为10至100微米的小通道中工作得最好。SPR对金传感垫周围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的混合物中快速分离和浓缩分析物。高分辨率的分离是可能的，通过介电泳法或可寻址的电泳法捕获可以实现1000倍的预浓缩。

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.