Studies On The New Enhanced Chemiluminescence Enzyme-Linked Immunoassay Systems And Its Applications

This paper described two kinds of phenol derivates: 4-hydroxy-4'-iodobiphenyl(HIOP) and 4-(1,2,4-triazol-1-yl)phenol(TRP) which were used as novel chemiluminescent enhancers in the Luminol-HRP-H2O2 chemiluminescene system and their applications of new enhanced chemiluminescene enzyme-linked immunoassay system. The results showed that these new methods are of high sensitivity, rapidity and simplicity, and could be used in the field of biological analysis. The main jobs of this thesis can be summarized as follows:First, a novel para-phenol derivative 4-(1,2,4-triazol-1-yl)phenol(TRP) was used as a highly potent enhancer in the new luminol-H2O2-horseradish peroxidase-HRP enhancend chemiluminescence system and optimized the influential factors such as the concentration of luminal, enhancement and H2O2; and the effect of buffer solution. The CL reaction conditions and its enhancement characteristics were optimized and compared with that of 4-iodophenol. Under the optimum conditions, the TRP was detected with a linear of 2.0×10-9 ~4×10-8 g/mL and a detection limit of 5.0×10-10 g/mL. The chemiluminescent enhancement effect of the TRP was good and can be used as biosensor for the determination of hydrogen peroxide.Second, this paper established a new enhancend chemiluminescence system used HIOP as a highly potent enhancer in the luminol-H2O2-horseradish peroxidase. The influential factors such as the time, the concentration and solution were optimized. Under the optimum conditions, the HIOP was detected with a linear of 5.0×10-11 ~4×10-8 g/mL and a detection limit of 9.0×10-12 g/mL, which was 10 folds lower than that of classical luminol-H2O2-horseradish peroxidase method. It can be concluded that the new system could be applied in the tumor markers. And the light emission mechanism of the new system was studied.Third, a fast and sensitive CL immunoassay method: Luminol-HRP-H2O2-HIOP was developed and used in detecting theα-AFP with the magnetic beads. Theα-AFP was detected by this method with the linear range of 1.0~150 ng/mL, and the correlation coefficient was 0.9993 and the detection limit of 0.5 ng/mL, which was 10 folds lower than that of general ELISA method. Under the optimum conditions,α-AFP samples were detected and the results showed good corresponding relationship.Four, the CEA antigen was detected by this new enhanced CL immunoassay method with the linear range of 0.5~40 ng/mL, and the correlation coefficient was 0.9989 and the detection limit of 0.5 ng/mL, which was 10 folds lower than that of general ELISA method. And the results of this new methods used in detecting the human blood serum showed good corresponding relationship.