Determination of Molecular Weights in Polyelectrolyte Mixtures Using Polymer Translocation through a Protein Nanopore

[Image: see text] We introduce a single molecular analysis technique for the evaluation of molecular weight distributions of polyelectrolyte solutions by measuring translocation times of sodium polystyrenesulfonate (NaPSS) chains in a mixture passing through an α-hemolysin protein nanopore. The ionic current through an α-hemolysin nanopore is partially blocked transiently when the pore is occupied by a polymer chain with an average residence time proportional to the molecular weight of the polymer chain. We have measured the translocation times for an equimolar mixture of four different molecular weight NaPSS standards and observed distinct translocation time distribution peaks, each of which corresponding to the different components in the mixture. Size exclusion chromatography analyses were performed on the equimolar and equiweight NaPSS mixtures of the same components and compared with the translocation time measurements. The experimental results demonstrate that measuring translocation times can be a competitive technique for estimating the broad molecular weight distributions of polyelectrolytes.