Electrochemical immunosensor of tumor necrosis factor α based on alkaline phosphatase functionalized nanospheres

A novel immunosensor for sensitive detection of tumor necrosis factor α was reported. First of all, gold nanoparticles were uniformly assembled on the surface of poly (styrene-acrylic acid) nanospheres, which was used as the matrix to conjugate alkaline phosphatase (ALP). And then, the obtained composite was used as multi-enzyme functionalized label for immunoassay. Biocompatible polyaniline doped with poly (acrylic acid) was electro-polymerized at the glass carbon electrode to construct the matrix for the immobilization of antibody TNF-α. After the sandwich immunoreaction, the labeled ALP was used to hydrolyze α-naphthyl phosphate to produce the electroactive α-naphthol, which could be amperometrically detected. The results showed that the electrochemical signals were proportional to the logarithm of the antigen concentration in the range of 0.02–200.00 ng/mL with the detection limit of 0.01 ng/mL. The developed immunoassay showed high sensitivity, acceptable stability and reproducibility, which might have potentially broad applications in protein diagnostics and bioassay.