Easy-to-Engineer Flexible Nanoelectrode Sensor from an Inexpensive Overhead Projector Sheet for Sweat Neuropeptide-Y Detection

In this paper, we report an inexpensive and easy-to-engineer flexible nanobiosensor electrode platform by exploring a nonconductive overhead projector (OHP) sheet for sweat Neuropeptide-Y (NPY) detection, a potential biomarker for stress, cardiovascular regulation, appetite, etc. We converted a nonconductive OHP sheet into a conductive nanobiosensor electrode platform with a hybrid polymerization method, which consists of interfacial polymerization of pyrrole and a template-free electropolymerization technique to decorate the electrode platform with poly(EDOT-COOH-co-EDOT-EG3) nanotubes. The selection of poly(EDOT-COOH) features an easy conjugation of NPY antibody (NPY-Ab) through EDC/Sulfo-NHS coupling chemistry, while poly(EDOT-EG3) is best known to reduce nonspecific binding of biomolecules. The antibody conjugation on the polymer surface was characterized by a quartz crystal microbalance, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, and chronoamperometry techniques. The OHP nanosensor platform exhibited the successful detection of NPY analyte through a chronoamperometry method in phosphate-buffered saline with a wide range of concentrations from 1 pg/mL to 1 μg/mL with a limit of detection of 0.68 pg/mL having good linearity (R2 = 0.9841). The sensor platform exhibited excellent stability, reproducibility, repeatability, and a shelf-life of 13 days. Furthermore, the sensor showed superior selectivity to a 100 pg/mL NPY analyte among other interfering compounds such as tumor necrosis factor α, cortisol, and Interleukin-6. The clinical practicality of the sensor was confirmed through the detection of 100 pg/mL NPY spiked artificial perspiration, highlighting the possibility of integrating the sensor platform to wearable healthcare applications.