Electrochemical Detection of Neuronal Injury in Cell Culture Samples: A Cost-Effective Biosensor for Neurofilament...

Neurofilament light chain (NfL) is a promising biomarker of axonal injury across acute and chronic neurodegeneration, which can improve drug discovery and disease monitoring models. Traditional in vivo animal models cannot fully mimic human pathophysiology of neurodegenerative diseases (NDDs), but in vitro models based on human cells solve this problem, reducing the time and cost of drug testing. We developed an electrochemical immunosensor for NfL detection in cell culture media to monitor acute neuronal injury in in vitro models. The biosensor was designed in two configurations: the label-free system, which directly detects NfL in the sample via the antibody-antigen interaction, and the sandwich configuration, which incorporates two additional antibodies. Detection was examined using electrochemical techniques, including cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and chronoamperometry (CA). The sensor demonstrated a detection limit of 3-9 pg mL^-1, and a dynamic working range spanning from 10 up to 10⁷ pg mL^-1. Importantly, NfL was successfully detected in physiological media collected from cultured neurons that were differentiated from the long-term human neuroepithelial-like stem cells. This discovery highlights the platform's applicability for in vitro neurodegenerative models. The immunosensor offers a sensitive, scalable, and cost-effective alternative for neurodegeneration detection in drug testing applications.