Photoelectrochemical sandwich immunoassay of brain glycogen phosphorylase based on methyl orange–sensitized TiO2 nanorods

• Published in: Mikrochimica acta (1966) 2022-07, Vol.189 (7), p.265-265, Article 265
• Main Authors: Sun, Chenglong, Li, Lu, Liu, Jialin, Du, Yun, Peng, Yueyi, Xie, Qingji
• Format: Article
• Language: English
• Subjects: Amplification; Analytical Chemistry; Antibodies; Ascorbic acid; Cadmium sulfide; Characterization and Evaluation of Materials; Chemical synthesis; Chemistry; Chemistry and Materials Science; Dyes; Electrochemistry; Electrodes; Electron microscopy; Fluorine; Fourier transforms; Glycogens; Heterojunctions; Immunoassay; Infrared spectra; Infrared spectroscopy; Microengineering; Microscopy; Nanochemistry; Nanorods; Nanotechnology; Original Paper; Oxidation; Phosphorylases; Photoelectric effect; Photoelectric emission; Silicon dioxide; Spectrum analysis; Tin oxides; Titanium dioxide
• ISSN: 0026-3672; 1436-5073
• DOI: 10.1007/s00604-022-05367-6

The photoelectrochemical immunoassay of glycogen phosphorylase BB (GPBB) was studied. A methyl orange/TiO 2 nanorod heterojunction was constructed on a fluorine-doped tin oxide electrode by hydrothermal synthesis, calcination, and chemical adsorption. A sandwich immune structure consisting of GPBB as the first antibody, GPBB, and a CdS@mesoporous silica-ascorbic acid (AA)–GPBB as secondary antibody composite was constructed on each of the selected well surfaces of a 96-well microplate. By adding mercaptoethylamine to structurally destroy the secondary antibody composite and release the electron donor AA, the amplification of photocurrent, and thus the “off–on” photoelectrochemical biosensing of GPBB were realized. The use of the 96-well microplate provides good reproducibility of the assembled immune structures and eliminates the possible effect of the photogenerated hole–induced protein oxidation on the photocurrent. The relevant electrodes and materials were characterized by electrochemistry, UV–vis diffuse reflectance spectra, Fourier transform infrared spectroscopy, X-ray diffractometer, scanning electron microscopy/energy dispersive spectroscopy, transmission electron microscopy and BET method. Under the optimal conditions, the photocurrent was linear with the logarithm of GPBB concentration from 0.005 to 200 ng mL −1 and with a limit of detection of 1.7 pg mL −1 ( S / N  = 3). Satisfactory results were obtained in the analysis of real serum samples. Graphical abstract A sandwich immune structure consisting of GPBB first antibody, GPBB, and a CdS@mesoporous silica-ascorbic acid (AA)–GPBB secondary antibody composite was constructed on each of the selected well surfaces of a 96-well microplate. By adding mercaptoethylamine to structurally destroy the secondary antibody composite and release the electron donor AA, the amplification of photocurrent, and thus the “off–on” photoelectrochemical biosensing of GPBB were realized.