Systematic Approach to DNA Aptamer Design Using Amino Acid–Nucleic Acid Hybrids (ANHs) Targeting Thrombin

Chemical modifications of innate DNA/RNA aptamers facilitate the improvement their function. Herein, we report our modular strategy to manipulate a thrombin-binding DNA aptamer (TBA) improve its anticoagulation activity and binding affinity. A set amino acid conjugates, termed acid–nucleic hybrids or ANHs, was synthesized incorporated into TBA loop sequences. We found that substitutions with hydrophobic acids in region possessed significantly enhanced antithrombin activity, up 3-fold higher than native TBA. investigated correlations between affinity features amino-acid conjugates using experimental techniques including circular dichroism spectroscopy, surface plasmon resonance assay, molecular modeling. The present study demonstrates systematic approach design based on characteristics, allowing development advanced aptamers.