The need for a robust, specific and high-throughput biosensing platform is of significant interest in analysis of glycans and oligosaccharides. However, current analytical technology requires long run times, sophisticated and expensive equipment (e.g., HPLC-MS, HPAEC-PED). In particular, the carbohydrate linkage is especially difficult to discern. Paper based biosensors provide alternative approaches for rapid and easy-to-use assays. We have developed a <b>linkage-specific assay with colorimetric readout</b> visible to the naked eye or analyzed using imaging software for quantitative measurements that are a function of the concentration of the target molecule. <b>The assay takes approximately 30 seconds and is amenable to high throughput via multichannel pipetting. </b>Glycolytic enzymes are used alongside glycosyl-dehydrogenase to generate a redox system capable of producing a colorimetric change in organic dyes. Heterologous expression of enzymes and immobilization on paper substrate increases the commercial feasibility of the assay in terms of cost and durability. This paper based analytical device platform demonstrates its utility in qualitative and quantitative assays with potential for expansion to other molecules and applications. In this work, we demonstrate the paper-based quantitation of a1-2, a1-3, and a1-6 fucose in the context of human milk oligosaccharides, Lewis antigens, and core N-glycans. We also present a method for quantifying degree and linkage characteristics of sialylation.

Linkage-specific, paper-based assay for characterization of N-glycans, O-glycans, and oligosaccharides

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.