Lipid-coated zero-mode wave-guide (ZMW) nanopores for single-molecule investigations of biomolecules.

Single-molecule (SM) methods provide a powerful way to investigate e.g. transient subpopulations and the dynamics of enzymes, features that are usually obscured by ensemble averaging. The rapid expansion of these technologies is revolutionizing our ability to probe the mechanisms of biological reactions in biological systems of ever-increasing complexity. Technological improvements have made it possible to combine several different single-molecule methods, which have opened yet new possibilities to study complex biochemical reactions on the single-molecule level. In this project we aim for the development of zero-mode waveguides (ZMW) nanopores covered with lipid bilayers for the high-throughput and single-molecule analysis of membrane proteins, including biological nanopores, under a controllable applied potential. The ability of studying many proteins, but one at a time, will be an important feature of this platform. Membrane proteins constitute about 30% of the human proteins and represent 60% of the drug targets. ZMWs nanopores will provide a powerful technology for studying real-time dynamics of membrane proteins at physiological conditions of ligand concentrations and applied potentials. Furthermore, biological nanopores are used in (emerging) biotechnology applications, the most important of which is arguably single-molecule DNA sequencing. The technology described here will have applications in high-speed and high-throughput optical mapping of single DNA molecules.