Study On The Interaction Between Sulfamethoxazole And Serum Albumin By Surface Plasmon Resonance

The novel detection technology of surface plasmon resonance (SPR) assay is known to monitor interaction between bimoleculars real time and ascertain the kinds of reactants as well as their concentrate without isolation and purification. SPR sensors are valuable analytical tools with a broad range of applications from laboratory assays to study for food safety , drug discovery and environmental pollutant monitoring, which have the feature of high sensitivity, label free, small amount of sample and rapid speed of detection. The first surface plasmon resonance(SPR)sensor was developed in 1983 by Liedberg et al, since then, the research and application of SPR technology have shown extensive growth and gradually become the hot spot and research frontier in the biosensor field in the world. These characteristics made the SPR technique an easy, convenient and reliable one for determining the concentration and molecular weight, monitoring change in structure, measuring kinetic constant and binding specificity of individual biomolecules. For above reasons the SPR sensors grow as a new powerful technology in chemical and biological field.Along with raising of medical standards, the level of protein binding became an important factor. The content of this paper was based on SPR biosensor to detect interaction between drug and serum albumin. The main contents are listed as follows.1. The SPR biosensor was used to monitor directly the interaction between SMZ and BSA. Firstly, the basement membrane MPA , was fixed on the surface of the SPR gold surface by self-assembly. On the basement membrane, BSA was successively assembled. Using the series of concentration injection method of SMZ, the standard curve was constructed. This combination could be thought as the single molecule combination, which the combination could be considered as first-order reaction. According to the equation, we could obtain that KA=1.250×10~5.2. Based on the highly specific binding force between biotin and avidin, the bridge connection as well as the amplification, the biotin-avidin system was applied into SPR biosensor. Firstly, DCC method was exploited to synthesize the BNHS, of which the melting range was 210 to 213℃. In addition, the BNHS was characterized by IR spectroscopy and NMR spectroscopy, by which the BSA was marked. The basement membrane, the MPA, was fixed on the surface of the SPR gold surface by self-assembly. On the basement membrane, avidin and BSA were successively assembled, forming the biosensor with BSA on the surface. On this surface, SMZ can be combined and measured. With the concentration as X-axis and the response signal as Y-axis, a fit curve y=1.236×10~2 x-2.286 was obtained. Compared with the curve y=30.88x+0.7300 that about the direct action between SMZ and BSA, the signal of the former one was obviously strengthened and the linear relation magnified.3. The SPR biosensor was used to monitor directly the interaction between SMZ and HSA. Firstly, the basement membrane MPA, was fixed on the surface of the SPR gold surface by self-assembly. On the basement membrane, HSA were successively assembled. Using the series of concentration injection method of SMZ, the standard curve was constructed. This combination could be thought as the single molecule combination and the combining force of the molecules was weak, which the combination could be considered as first-order reaction. According to the equation, we could obtain that KA=1.044×10~5.4. The biotin-avidin system was applied into SPR biosensor. The BSA was marked with synthesized BNHS. The basement membrane, the MPA, was fixed on the surface of the SPR gold surface by self-assembly. On the basement membrane, avidin and BSA were successively assembled, forming the biosensor with BSA on the surface. On this surface, SMZ can be combined and measured. With the concentration as X-axis and the response signal as Y-axis, a fit curve y=y=1.267×10~2 x-3.750 was obtained. Compared with the curve y=37.76x-1.546 that about the direct action between SMZ and BSA, the signal of the former one was obviously strengthened and the linear relation magnified. The results indicate that signal and the detection accuracy can be strengthened for the introduction of biotin-avidin system.