Tetra-substituted amino aluminum phthalocyanine as a new red-region fluorescent substrate for horseradish peroxidase based enzyme-linked immunosorbent assay
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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 H2O2 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 x 10(-6) mol/L-1 and 6.0 x 10(-9) mol (.) L-1 . s(-1), respectively. In this paper, TAAlPc was used in an HRP-based enzyme-linked immunosorbent may (ELISA) of alpha-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 radio-immunoassay. The advantage of proposed method was strongly minimizing the interference resulting from background fluorescence or scattering light and had a high and typical sensitivity.