Multiplex glycan bead array for high throughput and high content analyses of glycan binding proteins

Sharad Purohit,Peng George Wang,Xuezheng Song,Wanyi Guan,Tiehai Li,Yanna Tian,Jing Song,Lei Li,Bunja Rungruang,Boying Dun,Sharad Ghamande,Jin-Xiong She,Richard D Cummings,David Mysona,Ashok Sharma

doi:10.1038/s41467-017-02747-y

Abstract

Glycan-binding proteins (GBPs) play critical roles in diverse cellular functions such as cell adhesion, signal transduction and immune response. Studies of the interaction between GBPs and glycans have been hampered by the availability of high throughput and high-content technologies. Here we report multiplex glycan bead array (MGBA) that allows simultaneous analyses of 384 samples and up to 500 glycans in a single assay. The specificity, sensitivity and reproducibility of MGBA are evaluated using 39 plant lectins, 13 recombinant anti-glycan antibodies, and mammalian GBPs. We demonstrate the utility of this platform by the analyses of natural anti-glycan IgM and IgG antibodies in 961 human serum samples and the discovery of anti-glycan antibody biomarkers for ovarian cancer. Our data indicate that the MGBA platform is particularly suited for large population-based studies that require the analyses of large numbers of samples and glycans.

Highlights

Glycan-binding proteins (GBPs) play critical roles in diverse cellular functions such as cell adhesion, signal transduction and immune response
Other GBPs include proteins any cell surface and secretory proteins produced by mammalian cells are modified by glycans, which play an essential role in the maintenance of diverse cellular functions
The ability to probe the interaction between glycans and GBPs is of critical importance to biomedical research and clinical care of patients

Summary

Introduction

Glycan-binding proteins (GBPs) play critical roles in diverse cellular functions such as cell adhesion, signal transduction and immune response. The three main mammalian GBP families are Ctype lectins, galectins and siglecs, which play critical roles in diverse cellular functions such as cell adhesion, signal transduction and immune response[6]. Other GBPs include proteins any cell surface and secretory proteins produced by mammalian cells are modified by glycans, which play an essential role in the maintenance of diverse cellular functions. Alterations to glycans, such as increased levels of truncation and branching as well as presence of unusual terminal sequences, may cause changes in various physiological and Glycan. In addition to blood transfusion, anti-glycan antibodies may have a tremendous potential as diagnostic and prognostic markers for other diseases such as cancer and autoimmunity[5,9,10,11]

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Conclusion