Study Of Enzyme Immobilization By Polystyrene Monolithic Column In Microfluidic Analytical System

Base on summarization enzyme reaction internal and overseas in micro-fluidic analytical system. Contrast the characteristics of a variety of micro-fluidic analytical system based enzyme reaction. A enzyme immobilization by polystyrene monolithic column in microfluidic analytical system was selected and the application of the system was researched in this paper.Polystyrene monolithic columns were prepared by in-situ copolymerization of Styrene. Divinylbenzene and AIBN were used as cross-linker and initiator, respectively in the reaction process. Moreover, Ethanol was used as porogenic reagents. In order to immobile enzyme in the polystyrene monolithic columns, then via modified with sulfonated group, sulfonated polystyrene monolithic column was obtained. Further, its framework and speciality was charactered by some methods such as scan electron microscope (SEM) and infrared spectrum (IR). When the spead lower 1.5mL/min, adsorption column capacity to L-Tyr of polystyrene monolithic micro-column (1mm i.d×2.0cm) was 3.4mg and column content was 0.43mg/mm3.In addition,the column has a good mechanical stability.GOD was fixed on polystyrene monolithic micro-column (1mm i.d×2.0cm).Used UV detection determined when the use of free enzyme for 10 U, GOD was fixed about 7 U, and a fixed-efficiency up to 70 percent.The polystyrene monolithic column was In-situ polymered and modified with sulfonated group in Micro-fluidic chip. GOD was fixed in the chip.The optimized HRP-determined speed and concentration were 0.2 mL/min and 5ug/mL, respectively. Combined with the monolithic column fixed enzyme and micro chip techniques, a microchip with enzyme reaction was constructed.The microchip with enzyme reaction combined chemiluminescence detection, constructed a enzyme immobilization by polystyrene monolithic column in microfluidic - chemiluminescence analytical method. The optimized detections were that the spead of sample was 0.2mL/min and the ratio of sample and Luminol was 1:1. The constructed detection method was used to detect Vc in the vegetable samples and levodopa in blood.The determination was good. In the constructed detection method, via immobiled different enzyme or antibody could be used to analysis Corresponding enzyme substrate or antigen. The constructed detection method has a wide range of practicality.