We used base-appended base modification to develop a new adenine analog, which incorporates an adenine derivative at position 7 of adenine. Using the systematic evolution of ligands by exponential enrichment method with a modified DNA library including this analog, we obtained A, an aptamer that binds strongly to human β-defensin 2, a biomarker of physical stress found in saliva. The dissociation constant of A with respect to human β-defensin 2 was found to be low (6.8 nM), and was found to bind specifically to human β-defensin 2 in saliva spiked with the protein, as confirmed using pull-down with magnetic beads. To our knowledge, there are no prior reports of nucleic-acid aptamers that bind specifically to human β-defensin 2. However, our results indicated that such adenine analog-containing DNA libraries are extremely effective in the acquisition of high-affinity aptamers.
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http://dx.doi.org/10.1016/j.ab.2020.113627 | DOI Listing |
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