Based On The Spr Sensor Chip Technology Developed And Its Application

The biosensor based on the principle of surface plasmon resonance (SPR), is a surface sensitive optical device for monitoring biomolecular interactions at the sensor surface in real time without any labeling. It is being used in a wide variety of areas such as proteomics, drug discovery, clinical diagnosis, food analysis, environmental monitoring and so on. The sensor chip is the core of the Biacore instrument. Since sensor chips can now only be purchased from Biacore AB (Uppsala, Sweden), which makes them expensive, many instruments are either used at a low rate or sleeping. Besides, few types of sensor chips and difficult immobilization of materials to the sensor chip surface, are hindering the application of SPR biosensor to a certian extent.In this paper, in order to reduce the cost of snesor chips, the preparation and regeneration methods of the J1, C1, CM5, SA and NTA sensor chip were studied, and their preparation process were perfected. Through substantial experimental verification as well as a number of research institutes' trials, the self-prepared sensor chip can achieve the same quality as the Biacore sensor chip and meet the requirements of the experiment. In order to increase the types of optional sensor chips, a reduced bovine serum albumin (rBSA) sensor chip was developed, which is a new sensor chip for the Biacore instrument and can achieve the same function as the CM5 sensor chip. The preparation of rBSA sensor chip is simple, rapid and low cost. The atomic force microscopy (AFM) characterization of the rBSA sensor chip surface and the application to detect sulfamethoxazole (SMX) by the rBSA sensor chip using an inhibitive immunoassay format, demonstrated homogeneous matrix surface and excellent performance of the rBSA sensor chip, which is able to meet the needs of the experiment and the requirement of quantitative analysis of SMX. In order to improve the binding performance of the NTA sensor chip, a higher molecular density of a new improved NTA sensor chip was developed. The preparation method is rapid, simple and efficient. The binding of seven kinds of 6×His-tag recombinant protein to both improved and non-improved NTA sensor chip, respectively, indicated that the improved NTA sensor chips have excellent properties.In addition, we used the self-prepared CM5 chips to simulate the stainless steel endovascular stent surface, and to evaluate the effects of Chitosan samples with various molecular weights (MW) on the protection of plasmid DNA from nuclease degradation by SPR. The results showed that SPR can be used as a new method to study the stability of gene binding and to screen new gene carrier materials. Subsequently, we proposed and took advantage of a new immunoassay method using SPR coupled with on-line, in-tube, solid-phase microextraction (SPME), to detect estradiol in human serum and seawater using an indirect inhibitive immunoassay format. This method not only substantially simplifies the analytical process and shortens analysis time, but it also provides greater accuracy and precision than can be obtained with the traditional direct inhibition immunoassay. To date, to the best of our knowledge, no in-tube SPME-SPR methodology applied to substance detection has been reported in the literature.To sum up, several preparation and regeneration methods of a variety of SPR sensor chips have been developed in this paper, which not only greatly reduces the equipment running cost and improves the analytical capacity of the SPR biosensor, but also lays the foundation for further application of the SPR biosensor and development of domestic SPR biosensor. In addition, through the application of self-prepared sensor chips to various biomolecular interaction analysis and substance detection, this work provides new possibilities for application of SPR biosensor in the new areas.