Nanopore-based 5D fingerprinting of single proteins in real-time (London Calling Presentation)

Are 1-D and 2-D gel-electrophoresis the best we can do in routine protein analysis? This talk demonstrates that by moving proteins through nanopores, it is possible to separate these proteins transiently from other macromolecules in solution. This unique feature has the potential to enable instantaneous and unambiguous characterization of the one protein in the pore. We show that monitoring the rotational dynamics of this protein inside the pore then makes it possible to determine several excellent protein descriptors simultaneously, including the protein’s volume, shape, charge, dipole moment, and rotational diffusion coefficient. The resulting 5-dimensional protein characterisation may open the door to real-time protein characterisation, identification, and quantification on a single molecule level in complex mixtures. We think that this approach has the potential to be more sensitive, specific, quantitative, reproducible, and rapid than gel-electrophoresis while providing additional protein descriptors and requiring only minute sample sizes. This technique may replace the ubiquitous use of gel-electrophoresis for protein analysis all around the world with a chip-based approach to instantaneous protein counting that may also allow single protein sorting.