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  1. null (Ed.)
    Nanopore probing of molecular level transport of proteins is strongly influenced by electrolyte type, concentration, and solution pH. As a result, electrolyte chemistry and applied voltage are critical for protein transport and impact, for example, capture rate ( C R ), transport mechanism ( i.e. , electrophoresis, electroosmosis or diffusion), and 3D conformation ( e.g. , chaotropic vs. kosmotropic effects). In this study, we explored these using 0.5–4 M LiCl and KCl electrolytes with holo-human serum transferrin (hSTf) protein as the model protein in both low (±50 mV) and high (±400 mV) electric field regimes. Unlike in KCl, where events were purely electrophoretic, the transport in LiCl transitioned from electrophoretic to electroosmotic with decreasing salt concentration while intermediate concentrations ( i.e. , 2 M and 2.5 M) were influenced by diffusion. Segregating diffusion-limited capture rate ( R diff ) into electrophoretic ( R diff,EP ) and electroosmotic ( R diff,EO ) components provided an approach to calculate the zeta-potential of hSTf ( ζ hSTf ) with the aid of C R and zeta potential of the nanopore surface ( ζ pore ) with ( ζ pore – ζ hSTf ) governing the transport mechanism. Scrutinization of the conventional excluded volume model revealed its shortcomings in capturing surface contributions and a new model was then developed to fit the translocation characteristics of proteins. 
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  2. Abstract

    Electrolyte chemistry plays an important role in the transport properties of analytes through nanopores. Here, we report the translocation properties of the protein human serum transferrin (hSTf) in asymmetric LiCl salt concentrations with either positive (Ctrans/Ccis< 1) or negative chemical gradients (Ctrans/Ccis> 1). Thecisside concentration was fixed at 4 M for positive chemical gradients and at 0.5 M LiCl for negative chemical gradients, while thetransside concentration varied between 0.5 to 4 M which resulted in six different configurations, respectively, for both positive and negative gradient types. For positive chemical gradient conditions, translocations were observed in all six configurations for at least one voltage polarity whereas with negative gradient conditions, dead concentrations where no events at either polarity were observed. The flux of Li+and Clions and their resultant cation or anion enrichment zones, as well as the interplay of electrophoretic and electroosmotic transport directions, would determine whether hSTf can traverse across the pore.

     
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  3. Abstract

    Recently, we developed a fabrication method—chemically‐tuned controlled dielectric breakdown (CT‐CDB)—that produces nanopores (through thin silicon nitride membranes) surpassing legacy drawbacks associated with solid‐state nanopores (SSNs). However, the noise characteristics of CT‐CDB nanopores are largely unexplored. In this work, we investigated the 1/fnoise of CT‐CDB nanopores of varying solution pH, electrolyte type, electrolyte concentration, applied voltage, and pore diameter. Our findings indicate that the bulk Hooge parameter (αb) is about an order of magnitude greater than SSNs fabricated by transmission electron microscopy (TEM) while the surface Hooge parameter (αs) is ∼3 order magnitude greater. Theαsof CT‐CDB nanopores was ∼5 orders of magnitude greater than theirαb, which suggests that the surface contribution plays a dominant role in 1/fnoise. Experiments with DNA exhibited increasing capture rates with pH up to pH ∼8 followed by a drop at pH ∼9 perhaps due to the onset of electroosmotic force acting against the electrophoretic force. The1/fnoise was also measured for several electrolytes and LiCl was found to outperform NaCl, KCl, RbCl, and CsCl. The 1/fnoise was found to increase with the increasing electrolyte concentration and pore diameter. Taken together, the findings of this work suggest the pH approximate 7–8 range to be optimal for DNA sensing with CT‐CDB nanopores.

     
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  4. Abstract

    Polysaccharides have key biological functions and can be harnessed for therapeutic roles, such as the anticoagulant heparin. Their complexity—e.g., >100 monosaccharides with variety in linkage and branching structure—significantly complicates analysis compared to other biopolymers such as DNA and proteins. More, and improved, analysis tools have been called for, and here we demonstrate that solid-state silicon nitride nanopore sensors and tuned sensing conditions can be used to reliably detect native polysaccharides and enzymatic digestion products, differentiate between different polysaccharides in straightforward assays, provide new experimental insights into nanopore electrokinetics, and uncover polysaccharide properties. We show that nanopore sensing allows us to easily differentiate between a clinical heparin sample and one spiked with the contaminant that caused deaths in 2008 when its presence went undetected by conventional assays. The work reported here lays a foundation to further explore polysaccharide characterization and develop assays using thin-film solid-state nanopore sensors.

     
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