Optimizing the sensitivity and resolution of hyaluronan analysis with solid-state nanopores

Abstract

Hyaluronan (HA) is an essential carbohydrate in vertebrates that is a potentially robust bioindicator due to its critical roles in diverse physiological functions in health and disease. The intricate size-dependent function that exists for HA and its low abundance in most biological fluids have highlighted the need for sensitive technologies to provide accurate and quantitative assessments of polysaccharide molecular weight and concentration. We have demonstrated that solid state (SS-) nanopore technology can be exploited for this purpose, given its molecular sensitivity and analytical capacity, but there remains a need to further understand the impacts of experimental variables on the SS-nanopore signal for optimal interpretation of results. Here, we use model quasi-monodisperse HA polymers to determine the dependence of HA signal characteristics on a range of SS-nanopore measurement conditions, including applied voltage, pore diameter, and ionic buffer asymmetry. Our results identify important factors for improving the signal-to-noise ratio, resolution, and sensitivity of HA analysis with SS-nanopores.

Elucidating the Dynamics of Polymer Transport through Nanopores using Asymmetric Salt Concentrations

“…The most likely primary source of translocation time modulation is therefore the local concentration inside the pore modulating the electrophoretic mobility of dsDNA, for which slower translocations are expected in higher salt concentrations [46,47]. This is further supported by the dependence of translocation time velocity 𝓋 on salt gradient Ctrans/Ccis being well described by a decaying exponential (fit shown in Figure 5a inset), a dependence previously reported by Rivas et al [47].…”

“…have revealed both increased capture rates and decreased translocation velocities [37,[39][40][41][42][43][44][45], whereas these quantities are positively correlated when modulated by other standard means (e.g. tuning the voltage, salt concentration [46,47], pore size [36,47] or temperature [48]).…”

Glycosaminoglycans: What Remains To Be Deciphered?

“…Gas-phase electrophoretic mobility molecular analysis (GEMMA) and, more recently, solid-state nanopore sensors , are well suited to quantify HA mass. These techniques analyze one molecule at a time and thus provide the mass distribution of HA samples with exquisite detail beyond what is encompassed by the average mass and mass dispersity values.…”

Application of Nanopore Single Molecule Detection Technology in Analysis of Xylan Dissolved in Ionic Liquid†

“…[ 15 ] Especially for charged glycosaminoglycans and other water‐soluble polysaccharides, the molecular weight distribution of polysaccharides and molecular differentiation of polysaccharides with different modified groups have been achieved, but less attention has been paid to the detection of water insoluble polysaccharides. [ 16‐22 ] In this study, beech xylan (BX) extracted from beech was used as the research object of homogeneous xylan, and arabinoxylan (AX) extracted from corncob was used as the research object of heterogeneous xylan. The structures of AX and BX were shown in Figure S1.…”