Journal article
The Protein-Templated Synthesis of Enzyme-Generated Aptamers
Biologically Inspired Material Engineering, Biomimetics, Department of Health Technology, Technical University of Denmark1
Biomimetics, Department of Health Technology, Technical University of Denmark2
Department of Health Technology, Technical University of Denmark3
AI for Immunological Molecules, Bioinformatics, Department of Health Technology, Technical University of Denmark4
Bioinformatics, Department of Health Technology, Technical University of Denmark5
Technical University of Denmark6
International Iberian Nanotechnology Laboratory7
Biotherapeutic Engineering and Drug Targeting, Department of Health Technology, Technical University of Denmark8
Nanomaterials and Nanobiosensors, Biotherapeutic Engineering and Drug Targeting, Department of Health Technology, Technical University of Denmark9
Inspired by the chemical synthesis of molecularly imprinted polymers, we demonstrated for the first time, the protein-target mediated synthesis of enzyme-generated aptamers (EGAs). We prepared pre-polymerisation mixtures containing different ratios of nucleotides, an initiator sequence and protein template and incubated each mixture with terminal deoxynucleotidyl transferase (TdT).
Upon purification and rebinding of the EGAs against the target, we observed an enhancement in binding of templated-EGAs against the target compared to a non-templated control. These results demonstrate the presence of two primary mechanisms for the formation of EGAs, namely, the binding of random sequences to the target as observed in systematic evolution of ligands by exponential enrichment (SELEX) and the dynamic competition between TdT enzyme and the target protein for binding of EGAs during synthesis.
The latter mechanism serves to increase the stringency of EGA-based screening and represents a new way to develop aptamers that relies on rational design.
Language: | English |
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Year: | 2022 |
Pages: | e202201061 |
ISSN: | 15213773 and 14337851 |
Types: | Journal article |
DOI: | 10.1002/anie.202201061 |
ORCIDs: | 0000-0003-2428-6835 , Marcatili, Paolo , Sun, Yi and Ashley, Jon |