Porphyrin Probe Capillary Electrophoresis Analysis Of Protein Method

The goal of this dissertation is to develop novel quantitative methods for protein with Capillary Electrophoresis(CE). The application of meso-tetrakis(4-sulfonatophenyl) Porphyrin(TPPS4) as probe to detect proteins by CE was studied and discussed the possibility of using the two kinds of porphyrins as tracer in Capillary Electrophoresis Immunoassays was discussed. The reaction of TPPS4 with protein was studied by UV-Vis spectrophotometry. The relationship between the concentration of protein and the UV-Vis absorption of TPPS4 solution at 435nm was reported. The related parameters such as reaction time, temperature, pH of the buffer, and concentration of TPPS4 on the quantitative were studied. The result show that BSA can be use as standard protein when albumin contains in protein samples more than 60%.With TPPS4 as tracer in CE, a sensitive method for the determination of protein was studied. The influence of changing the pH of buffer and adding (-CD after the reaction on the compound stability were discussed. Optimized the condition such as pH of buffer and type of buffers for electrophoresis were acquired. Adding (-CD to buffer can reduce the assay time and improve the shape of gauss peak of TPPS4 in capillary electrophoresis assay. This method can be used to detect protein in the range of 1-20(g/mL at correlation coefficients better than 0.993. The results showed that it is satisfactory to apply this method in the detection of proteins in human serum samples.The feasibility of using TPPS4 and hemin as tracer in immunoassays and CEIA were evaluated. The combinations of TPPS4 and hemin with protein were discussed. The combination rate of TPPS4-BSA and the affection on immunology activity was measured. The result indicate that the sensitiveness have no improvement when the protein marked with high sensitive dye porphyrin in CEIA with absorbance detection.