A Host Receptor Nanodisc-Based Biosensor Platform for the Detection of a Specific Virus and Its Variants

Abstract

The host receptor is a key element in the initial stage of the virus entry into the host. The use of this host receptor is valuable as a sensing element for selectively and sensitively detecting specific viruses. Also, viruses tend to escape neutralizing antibodies through viral mutation but still utilize the cell entry process using the same host receptors, so it would be a powerful detection tool even for the mutant viruses. The angiotensin-converting enzyme 2 (ACE2) receptor, which is the representative host receptor, performs an essential function in facilitating viral penetration by interacting with the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. In this study, we introduce a novel approach, where we fabricated a carbon nanotube field-effect transistor (CNT-FET) sensor and combined it with ACE2 receptor-embedded nanodisc (ND). ACE2 was produced using an E. coli expression system, purified, and integrated into the ND platform. ACE2 NDs showed robust functionality through interactions with a pseudotyped virus (PV) containing the spike protein, enabling sensitive detection of both SARS-CoV-2 and its genetic variations at 10(2) PFU/mL. The ACE ND-based sensor exhibited excellent selectivity by accurately differentiating SARS-CoV-2 wild-type and variants (Omicron, Delta) from other viruses (ZIKA and MERS-CoV). As a result of comparative analysis, ACE2 ND showed approximately 49% superior long-term functionality up to the second week compared to that of soluble ACE2. These findings highlight the high selectivity and sensitivity of host receptor-based sensors for detecting viral variants and provide a promising tool to prevent the spread of unknown viruses.