A neoglycoprotein-immobilized fluorescent magnetic bead suspension multiplex array for galectin-binding studies

Libo Zhang, Hai Yu, Yuanyuan Bai, Bijoyananda Mishra, Xiaoxiao Yang, Jing Wang, Evan B. Yu, Riyao Li, Xi Chen

Research output: Contribution to journalArticlepeer-review

Abstract

Carbohydrate-protein conjugates have diverse applications. They have been used clinically as vaccines against bacterial infection and have been developed for high-throughput assays to elucidate the ligand specificities of glycan-binding proteins (GBPs) and antibodies. Here, we report an effective process that combines highly efficient chemoenzymatic synthesis of carbohydrates, production of carbohydrate-bovine serum albumin (glycan-BSA) conjugates using a squarate linker, and convenient immobilization of the resulting neoglycoproteins on carboxylate-coated fluorescent magnetic beads for the development of a suspension multiplex array platform. A glycan-BSA-bead array containing BSA and 50 glycan-BSA conjugates with tuned glycan valency was generated. The binding profiles of six plant lectins with binding preference towards Gal and/or GalNAc, as well as human galectin-3 and galectin-8, were readily obtained. Our results provide useful information to understand the multivalent glycan-binding properties of human galectins. The neoglycoprotein-immobilized fluorescent magnetic bead suspension multiplex array is a robust and flexible platform for rapid analysis of glycan and GBP interactions and will find broad applications.

Original languageEnglish (US)
Article number6194
JournalMolecules
Volume26
Issue number20
DOIs
StatePublished - Oct 1 2021

Keywords

  • carbohydrate-protein conjugate
  • Galectin
  • Glycan-binding protein
  • Lectin
  • Multiplex assay

ASJC Scopus subject areas

  • Analytical Chemistry
  • Chemistry (miscellaneous)
  • Molecular Medicine
  • Pharmaceutical Science
  • Drug Discovery
  • Physical and Theoretical Chemistry
  • Organic Chemistry

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