Automated Label-Free Assay for Viral Detection and Inhibitor Screening via Biomembrane-Functionalized Microelectrode Arrays.

Abstract

Most virus infection assays have indirect readout such as virus number following entry (e.g., PCR, cell lysis). While effective, these technologies are labor-intensive, require specialized environments (e.g., sterile or RNA-free), and detect later-stage viral events like lysis or cell death, lacking sensitivity to early fusion events. To address these limitations, we present biologically relevant 2D membrane materials, host-cell-derived supported lipid bilayers (hcd-SLBs), integrated with organic microelectrode arrays (OMEAs) for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) fusion. By overexpressing angiotensin-converting enzyme 2 (ACE2) receptors on the native membranes, the platform functions as a viral sensor capable of detecting virus pseudo particles (VPPs) through the late pathway. Additionally, hcd-SLBs extracted from human lung epithelium expressing native ACE2 detect fusion events through the early pathway. The platform's utility as a drug-screening tool is demonstrated by testing antibodies targeting either the ACE2 on the host membrane or the viral spike (S) proteins. To enhance the throughput, microfluidics are integrated for automation and OMEAs are incorporated within each channel, miniaturizing the testing units. This system supports high-throughput data generation, automation, and scalability, providing an efficient platform for viral fusion detection that advances the study of pathogen-host interactions and accelerates antiviral drug discovery.