Antibody Immobilization in Zinc Oxide Thin Films as an Easy-Handle Strategy for Escherichia coli Detection

The antibody immobilization compatible with low-cost materials and label-free strategies is a challenge for biosensor device fabrication. In this study, ZnO thin film deposition was carried out on corning glass substrates by ultrasonic spray pyrolysis at 200 °C. The thin films were analyzed as platforms for enteropathogenic Escherichia coli (E. coli EPEC) antibody immobilization. The modification of thin films from the functionalization and antibody immobilization steps was visualized using Fourier transform infrared spectroscopy (FTIR) spectroscopy, and surface changes were observed by atomic force microscopy. The obtained FTIR spectra after functionalization showed a contribution of the amino group (NH₂) derived from silane (3-aminopropyltrimethoxysilane). The antibody immobilization showed an amide I conserved signal corresponding to the C=O stretching vibrations and the amide II signal related to the N-H scissor vibration mode. In this way, the signals observed are correlated with the presence of antibody immobilized on the film. The ZnO film morphology changes after every stage of the process and allows observing the antibody distribution on the immobilized surface. In order to validate the antibody recognition capability as well as the E. coli EPEC detection in situ, polymerase chain reaction was used.