Sensing and characterization of water-soluble peptides is of critical importance in a wide variety of bioapplications. Single molecule nanopore spectrometry (SMNS) is based on the idea that one can use biological protein nanopores to resolve different sized molecules down to limits set by the blockade duration and noise. Previous work has shown that this enables discrimination between polyethylene glycol (PEG) molecules that differ by a single monomer unit. This paper describes efforts to extend SMNS to a variety of biologically relevant, water-soluble peptides. We describe the use of Au(SG) clusters, previously shown to improve PEG detection, to increase the on- and off-rate of peptides to the pore. In addition, we study the role that fluctuations play in the single molecule nanopore spectrometry (SMNS) methodology and show that modifying solution conditions to increase peptide flexibility (via pH or chaotropic salt) leads to a nearly 2-fold reduction in the current blockade fluctuations and a corresponding narrowing of the peaks in the blockade distributions. Finally, a model is presented that connects the current blockade depths to the mass of the peptides, which shows that our enhanced SMNS detection improves the mass resolution of the nanopore sensor more than 2-fold for the largest cationic peptides studied.

The redox potential of packed red blood cells (RBCs) was measured over a 56-day storage period using a newly developed potentiometric methodology consisting of a nanoporous gold electrode and a silver chloride coated silver reference electrode. Both milliliter- and microliter-sized volumes were separately evaluated. The addition of Vitamin C (VitC) in differing doses to the packed RBCs was also assessed as a means to improve redox stability and prolong storage duration. For RBCs containing only saline, the open-circuit potential (OCP) was ∼ -80 mV vs Ag/AgCl and drifted slightly with time; greater differences were also noted between different electrodes. The addition of exogenous VitC to the RBC shifts the OCP to more negative values, stabilizes the redox potential, and improves reproducibly between different electrodes due to the poising of blood. Over the 56-day storage period, the redox potential of the RBCs increased slightly, which can be attributed to change in pH and/or increasing oxidative stress during storage. Cyclic voltammograms acquired after open-circuit potential measurements showed a characteristic peak attributed to the oxidation of VitC. This peak decreased during storage with a time constant of 20.8 days. Likewise, the intercellular concentration of VitC increased with a time constant of 20.2 days as measured using a fluorescence assay. Collectively, these results demonstrate the usefulness of electrochemical measurements in the study of stored blood products.