Cell Signaling Conduction And Communication Mechanism For Detection Of Glucagon And Cortisol

This thesis consists of three parts:In chapter one, an method for determination of glucose was developed based on double enzymes reaction. In this method , we used a solution containing glucose oxidase, horseradish peroxidase and 10-Acetyl-3,7-Dihydroxyphenoxazine. And we discussed the optimal conditions including excitation wavelength, concentration of GOD and HRP, the time of incubation. In the end of this chapter, we determined the calibration curve, the line scope and the limit of the detection of the double enzymes reaction under the optimal conditions.In chapter two, we investigated the cell sensor of glucagon based on the cell signaling transduction. The principle is: After glucagon combined with special receptor on the cell surface, the G protein within cells causes the signaling transduction to generate glucose. The generated glucose is released from the cell into the solution. The glucose in the solution is then detected using the method described in chapter one. The amount of glucagon can be measured through determining glucose concentration .Since the glucose concentration is higher than the glucagon concentration, implying amplification of the signal. It was found that the signal was amplified SO times.In chapter three, we detected the hydrocortison based on the theory of cell communication and signal transduction. Under the optimal conditions of the experiment, the limit of detection of the glucagon was 7.0×10^-9 mol/L and the linear scope was between 1.0×10^-5-2.3×10^-8 mol/L; the limit of detection of the hydrocortison was 1.4×10^-10 mol/L and the linear scope was between 8.0×10^-9 -5.0×10^-10 mol/L. It was found that the signal was amplified 134 times.