A solid-state nanopore-based single-molecule approach for label-free characterization of plant polysaccharides

Polysaccharides are important biomacromolecules existing in all plants, most of which are integrated into a fibrillar structure called the cell wall. In the absence of an effective methodology for polysaccharide analysis that arises from compositional heterogeneity and structural flexibility, our knowledge of cell wall architecture and function is greatly constrained. Here, we develop a single-molecule approach for identifying plant polysaccharides with acetylated modification levels. We designed a solid-state nanopore sensor supported by a free-standing SiNx membrane in fluidic cells. This device was able to detect cell wall polysaccharide xylans at concentrations as low as 5 ng/μL and discriminate xylans with hyperacetylated and unacetylated modifications. We further demonstrated the capability of this method in distinguishing arabinoxylan and glucuronoxylan in monocot and dicot plants. Combining the data for categorizing polysaccharide mixtures, our study establishes a single-molecule platform for polysaccharide analysis, opening a new avenue for understanding cell wall structures, and expanding polysaccharide applications. A solid-state nanopore sensor was developed to identify plant polysaccharides, which facilitates a single-molecular way to analyze such biomacromolecules. The established approach can effectively discriminate the most abundant noncellulosic cell wall polysaccharide xylans with varied acetylation levels and substituent patterns. Hence, this study offers a potential analytical new tool for polysaccharide characterization.