Lipid Bilayer Coated Nanopipettes as Generic Nanopore Sensors with Enhanced Functionality

Abstract

Nanopore sensors show great promise for use as single-molecule diagnostic devices. Current limitations to nanopores include laborious fabrication, undesired interactions between species in solution and the nanopore walls, pore clogging and difficulties in controlling the speed of translocations.Work in the literature shows the viability of using lipid coated solid-state pores and nanopipettes to overcome some of the aforementioned limitations to the development of nanopore biosensors.1,2 Such coatings offer increased control over the surface charge of the pore and the translocation speed, preconcentration of analytes on the pore surface prior to analysis, enhanced specificity and a reduction in pore clogging.Our work revolves around developing lipid-bilayer modified pores as a generic platform to detect epigenetic modifications of DNA. Compared to pull-down assays, the combination with nanopore analysis provides enhanced information content, while being faster and cheaper than single-base resolution bisulfite sequencing. Our data further show that the coated nanopipettes remain stable for hours and suitable for DNA translocation experiments. DNA methylation is being investigated as a model system, with particular relevance as a biomarker for early cancer diagnosis.1. Yusko, E. C. et al. Controlling protein translocation through nanopores with bio-inspired fluid walls. Nat. Nanotechnol. 6, 253-260 (2011).2. Hernandez-Ainsa, S. et al. Lipid-coated nanocapillaries for DNA sensing. Analyst 138, 104-106 (2013).