A new optical platform for biosensing based on fluorescence anisotropy
A new optical platform for biosensing based on fluorescence anisotropy
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DOI：
10.1007/s00216-008-1904-4
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发表时间：
2008-07-01
影响因子：
4.3
通讯作者：
Trono, C.
中科院分区：
文献类型：
--
作者：
Baldini, F.;Carloni, A.;Trono, C.
设计并研制了一种用于光学生物芯片检测的新型荧光光学平台。光学生物芯片由聚（甲基丙烯酸甲酯）（PMMA）制成，该PMMA由两片适当成形的PMMA形成，以获得四个500 μ m宽和400 μ m高的微通道。下部包括微通道和流体的入口和出口，而传感生物层固定在上部。光学信号包括由生物层发射的荧光，其各向异性地耦合到PMMA盖并由PMMA芯片适当地引导。通过在流动通道内进行的直接IgG/抗IgG相互作用，研究了光学芯片作为生物传感器的潜力。将小鼠IgG共价固定在PMMA盖的内壁上，并将Cy 5标记的抗小鼠IgG用于特异性相互作用。研究了PMMA表面的几种化学处理，聚（L-乳酸）、Eudragit L100和NaOH，以获得可用于共价固定小鼠IgG的羧基的最有效分布。发现用Eudragit L100处理是最成功的。采用所述配置，检测限和定量限分别为0.05 μ g mL（-1）和0.2 μ g mL（-1）。
A novel fluorescence-based optical platform for the interrogation of an optical biochip was designed and developed. The optical biochip was made of poly(methyl methacrylate) (PMMA) formed by two pieces of PMMA appropriately shaped in order to obtain four microchannels that are 500-mu m wide and 400-mu m high. The lower part includes the microchannels and the inlet and outlet for the fluidics, while the sensing biolayer was immobilized on the upper part. The optical signal comprised the fluorescence emitted by the biolayer, which was anisotropically coupled to the PMMA cover and suitably guided by the PMMA chip. The potentiality of the optical chip as a biosensor was investigated by means of a direct IgG/anti-IgG interaction carried out inside the flow channels. The mouse-IgG was covalently immobilized on the internal wall of the PMMA cover, and the Cy5-labelled anti-mouse IgG was used for the specific interaction. Several chemical treatments of the PMMA surface were investigated, poly(L-lactic acid), Eudragit L100 and NaOH, in order to obtain the most effective distribution of carboxylic groups useful for the covalent immobilisation of the mouse-IgG. The treatment with Eudragit L100 was found to be the most successful. Limits of detection and quantification of 0.05 mu g mL(-1) and 0.2 mu g mL(-1), respectively, were obtained with the configuration described.