The Protein‐Templated Synthesis of Enzyme‐Generated Aptamers

Inspired by the chemical synthesis of molecularly imprinted polymers, we demonstrated for first time, protein-target mediated enzyme-generated aptamers (EGAs). We prepared pre-polymerisation mixtures containing different ratios nucleotides, an initiator sequence and protein template incubated each mixture with terminal deoxynucleotidyl transferase (TdT). Upon purification rebinding EGAs against target, observed enhancement in binding templated-EGAs towards target compared to a non-templated control. These results demonstrate presence two primary mechanisms formation EGAs, namely, random sequences as systematic evolution ligands exponential enrichment (SELEX) dynamic competition between TdT enzyme during synthesis. The latter mechanism serves increase stringency EGA-based screening represents new way develop that relies on rational design.