Comparative Characterization of Diverse DNA Aptamers for Recognition of Spike Proteins of Multiple SARS‐CoV‐2 Variants

Many DNA aptamers have been reported to target the spike protein (S-protein) of SARS-CoV-2. These aptamers display different affinities or recognize different epitopes of the S-protein. We conducted a comparative study of 9 DNA aptamers for binding to several variants of the S-protein and pseudoviruses using the same testing methods, including dot-blot assays and enzyme-linked aptamer binding assays, to evaluate their affinity ranking and analytical utility. Moreover, the binding sites of these aptamers on the S-protein were examined using aptamer competition assays to understand the effect of S-protein mutations on aptamer affinity and the degree of overlapping of the binding sites by these aptamers. A head-to-head DNA aptamer comparison for SARS-CoV-2. DNA aptamers binding to several Variants of Concern (VOCs) of SARS-CoV-2 have been comparatively studied using the same methods to evaluate affinity ranking and guide future applications. The binding sites and overlapping degree of aptamers on the spike protein were examined to understand the effect of spike protein mutations on aptamer binding affinity.+image