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Generating enrichment through SELEX for perfluorobutanesulfonic acid and perfluoropentanoic acid-targeted aptamers
Bravo, Isabel A.
Bravo, Isabel A.
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2024
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Combinatorial approaches are leveraged to generate large random DNA oligomer libraries. Using SELEX (Systematic Evolution of Ligands by EXponential Enrichment), these libraries can be “panned” for sequences that bind a small molecule target with high affinity and selectivity. In this work, aptamers with two variable regions, one spanning 40 nucleotides and the other spanning 10 nucleotides, are immobilized and tethered to agarose beads via a constant region in the aptamer. Aptamers were challenged with short chain per- and polyfluoroalkyl substances (PFAS) and sequences with some affinity for the molecules were eluted from the beads and selected for subsequent rounds of enrichment. In total, twelve rounds of SELEX were performed. Significant enrichment began accumulating in the aptamer library starting as early as round six. Of the final 10 aptamer candidates, those present after twelve rounds of selection, four sequences were retained from round 6 and four were retained from round 9. These aptamers contain statistically significant sequence motifs, validating the design and application of SELEX. Further, the enriched motifs were positioned towards the outside of the variable regions, implicating the dissociation of the immobilizing region in short chain PF In future work, the final 10 aptamer candidates should be characterized for short chain PFAS binding affinity and selectivity.
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