Selection of optimised ligands by fluorescence-activated bead sorting.

ABSTRACT: The chemistry of aptamers is largely limited to natural nucleotides, and although modifications of nucleic acids can enhance target aptamer affinity, there has not yet been a technology for selecting the right modifications in the right locations out of the vast number of possibilities, because enzymatic amplification does not transmit sequence-specific modification information. Here we show the first method for the selection of specific nucleoside modifications that increase aptamer binding efficacy, using the oncoprotein EGFR as a model target. Using fluorescence-activated bead sorting (FABS), we have successfully selected optimized aptamers from a library of >65 000 variations. Hits were identified by tandem mass spectrometry and validated by using an EGFR binding assay and computationa

SUBMITTER: Paul AR 

PROVIDER: S-EPMC10498682 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature