Automated exo-glycosidase-based glycan sequencing on a digital microfluidic system with integrated electrochemical
Zhen Liu1, Riqiang Huang1, Mengyuan Zhao2
1Guangdong-Hong Kong-Macau Joint Laboratory on Micro-Nano Manufacturing Technology, Department of Materials Science and Engineering, Southern University of Science and Technology, Shenzhen, 518055, China.
Abstract:
Glycans are one of the most important biomacromolecules and are essential to nearly all known life forms. However, their structural complexity makes structural analysis more challenging than nucleic acids or proteins, significantly limiting the opportunities for investigating their structure-function relationship. Here, we proposed a novel glycan sequencing strategy in which a suite of specific non-reducing-end exoglycosidases was integrated with electrochemical detection of monosaccharides in a digital microfluidic (DMF) chip. In this study, a DMF platform with integrated non-enzymatic electrochemical sensors for monosaccharides was set up and characterized. Nine mammal monosaccharides were electrochemically investigated for the first time through cyclic voltammetry (CV) and pulse amperometry (PA) on this DMF platform. Automatic enzymatic hydrolysis of tagged lactoses was performed, and released monosaccharides were electrochemically detected and quantified, and the sequence of lactose was deduced. This current study would offer a promising new strategy for accurate and efficient sequencing of pure glycans with defined sequences and connectivity in their molecular structures.
