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The Protein-Templated Synthesis of Enzyme-Generated Aptamers
Angewandte Chemie Int. Ed. 2022
J.Vacacela, A.Schaap-Johansen, P. Manikova, P.Marcatili, M.Prado, Y.Sun, J.Ashley
This study demonstrated a protein-templated route to enzyme-generated aptamers (EGAs): nucleotide mixtures with an initiator sequence were extended by terminal deoxynucleotidyl transferase (TdT) in the presence of a protein target, producing aptamer pools with enhanced target binding versus non-templated controls — a rational-design alternative to conventional SELEX. Flow Induced Dispersion Analysis (FIDA), on a Fida 1 instrument, gave in-solution confirmation of aptamer–target binding, measuring binding affinity (KD, low-nanomolar) alongside EMSA and SPR. It is an example of using FIDA to measure aptamer–target binding affinity in solution.
