Print Email Facebook Twitter Orientation-Locked DNA Origami for Stable Trapping of Small Proteins in the Nanopore Electro-Osmotic Trap Title Orientation-Locked DNA Origami for Stable Trapping of Small Proteins in the Nanopore Electro-Osmotic Trap Author Wen, C. (TU Delft BN/Cees Dekker Lab; Wageningen University & Research; Kavli institute of nanoscience Delft) Bertosin, E. (TU Delft BN/Cees Dekker Lab; Kavli institute of nanoscience Delft) Shi, X. (TU Delft BN/Cees Dekker Lab; Kavli institute of nanoscience Delft) Dekker, C. (TU Delft BN/Cees Dekker Lab; Kavli institute of nanoscience Delft) Schmid, Sonja (Wageningen University & Research) Date 2022 Abstract Nanopores are versatile single-molecule sensors offering a simple label-free readout with great sensitivity. We recently introduced the nanopore electro-osmotic trap (NEOtrap) which can trap and sense single unmodified proteins for long times. The trapping is achieved by the electro-osmotic flow (EOF) generated from a DNA-origami sphere docked onto the pore, but thermal fluctuations of the origami limited the trapping of small proteins. Here, we use site-specific cholesterol functionalization of the origami sphere to firmly link it to the lipid-coated nanopore. We can lock the origami in either a vertical or horizontal orientation which strongly modulates the EOF. The optimized EOF greatly enhances the trapping capacity, yielding reduced noise, reduced measurement heterogeneity, an increased capture rate, and 100-fold extended observation times. We demonstrate the trapping of a variety of single proteins, including small ones down to 14 kDa. The cholesterol functionalization significantly expands the application range of the NEOtrap technology. Subject DNA origamielectro-osmotic flowlabel-free protein trappingnanopore electro-osmotic trap (NEOtrap)single-molecule detection To reference this document use: http://resolver.tudelft.nl/uuid:eeeb096c-2f7f-40c7-becd-28c064fcdce2 DOI https://doi.org/10.1021/acs.nanolett.2c03569 ISSN 1530-6984 Source Nano Letters: a journal dedicated to nanoscience and nanotechnology, 23 (3), 788-794 Part of collection Institutional Repository Document type journal article Rights © 2022 C. Wen, E. Bertosin, X. Shi, C. Dekker, Sonja Schmid Files PDF acs.nanolett.2c03569.pdf 4.4 MB Close viewer /islandora/object/uuid:eeeb096c-2f7f-40c7-becd-28c064fcdce2/datastream/OBJ/view