Molecular Crowding Modulates SARS-CoV-2 Aptamer Affinity

SARS-CoV-2 aptamer is a favorable candidate for the recognition and detection of SARS-CoV-2, owing to its small size and easy synthesis. However, the issue of compromised binding affinities in real samples and targeting mutant SARS-CoV-2 hinder wide applications of the aptamer. In this study, it is discovered that molecular crowding could increase binding affinity of CoV2-6C3 aptamer against RBD (Receptor Binding Domain) of SARS-CoV-2 via increasing the absolute value of the enthalpy change. The values of the equilibrium dissociation constant in molecular crowding decrease by 70% and 150%, respectively, against wild-type and mutant RBD compared with those in buffer without crowding. Moreover, the detection limit of SARS-CoV-2 pseudovirus is up to 5 times lower under molecular crowding compared to dilute conditions. The discovery deepens the understanding of aptamer-target interaction mechanisms in crowding conditions and provides an effective way to apply SARS-CoV-2 aptamer for virus recognition and detection.