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The use of tetra-substituted amino aluminum phthalocyanine (TAAlPc) as a new red-region fluorescent substrate for horseradish peroxidase (HRP)-based enzyme-linked immunosorbent assay was investigated. TAAlPc displayed an excitation maximum at 610 nm and emission maximum at 678 nm in a strong acidic medium. In the presence of HRP, trace amounts of H 2O 2 could rapidly and significantly react with TAAlPc, thus quenching the fluorescence of TAAlPc. The Michaelis-Menten parameters K m and V max were measured to be 2.82×10 -6 mol/L -1 and 6.0×10 -9 mol·L -1·s -1, respectively. In this paper, TAAlPc was used in an HRP-based enzyme-linked immunosorbent assay (ELISA) of α-fetoprotein (AFP) in human serum with satisfactory results. AFP could be determined in the concentration range of 0.5-200 ng/mL with a detection limit of 0.2 ng/mL, which was close to that of radioimmunoassay. The advantage of proposed method was strongly minimizing the interference resulting from background fluorescence or scattering light and had a high analytical sensitivity.
The use of tetra-substituted amino aluminum phthalocyanine (TAAlPc) as a new red-region fluorescent substrate for horseradish peroxidase (HRP) -based enzyme-linked immunosorbent assay was investigated. TAAlPc displayed an excitation maximum at 610 nm and emission maximum at 678 nm The Michaelis-Menten parameters K m and V max were measured to be 2.82 × 10 in the strong acidic medium. The presence of HRP, trace amounts of H 2 O 2 could rapidly and significantly react with TAAlPc, thus quenching the fluorescence of TAAlPc. -6 mol / L -1 and 6.0 × 10 -9 mol·L -1 · s -1, respectively. This paper, TAAlPc was used in an HRP-based enzyme-linked immunosorbent assay (ELISA) of α-fetoprotein AFP) in human serum with satisfactory results. AFP could be determined in the concentration range of 0.5-200 ng / mL with a detection limit of 0.2 ng / mL, which was close to that of radioimmunoassay. The advantage of proposed method was strongly minimizing the interference resulting from b ackground fluorescence or scattering light and had a high analytical sensitivity.