Study On The Detection Of Glycoprotein By Surface Plasmon Resonance Spectrometer

As a research focus in recent years, Surface Plasmon Resonance (SPR) technology, featured by real-time monitoring, label-free, no purification, high sensitivity, especially in the research of kinetic constant, has been widely applied to biochemical analysis, medical detection, environmental protection, food security and other fields.A surface plasmon resonance (SPR) biochemical analyzer is introduced in this paper. The method of combining angle scanning with intensity modulation is adopted to realize the real-time monitoring of glycoprotein. In the theory and simulation, first the principle of surface plasmon resonance is introduced; then simulation by Matlab theory illustrates the availability of scanning angle and intensity modulation method combing. The midpoint of the resonance angle and the slope maximum value is used the optimal operating point in the comparison of three different operating point, which is verified in the experimental section. We detect the glycoprotein in the experiment.In the experiment of system calibration, we verify the simulation conclusion, which is the best working point of midpoint. It is concluded that the system of linear range is 1.3256 to 1.36997, the sensitivity is 176.7RIU-1, the refractive index resolution is 1.248×10-5 RIU. In the experiments of glycoprotein detection, we choose the renewable liquid of pH=3, because the regenerative capacity of the liquid reaches 97% in the renewable solution of different pH value comparison; relative standard deviation (RSD) is 3.83% in the five measurements of the same concentration glycoprotein; our experimental method can detect the specificity of glycoprotein in the comparative experiments of non-glycoprotein and glycoprotein; finally, the glycoprotein concentration between 0.01mg/mL and 1mg/mL is detected, and polynomial fitting curve is used standard curve by comparing the residual analysis and correlation coefficients of linear fitting and polynomial fitting; the kinetic constant analysis obtains binding equilibrium constant KA=5.787mL/mg. The experimental results show that the SPR spectrometer can detect the specificity of glycoprotein, which will be extensively used in medical field.