High-Throughput Screening of Glycan-Binding Proteins Using Miniature Pig Kidney N-Glycan-Immobilized Beads

Academic Article

Abstract

  • Glycan recognition leading to cell-cell interactions, signaling, and immune responses is mediated by various glycan-binding proteins (GBPs) showing highly diverse ligand specificities. We describe here a rapid glycan immobilization technique via 4-hydrazinobenzoic acid (HBA)-functionalized beads and its application to high-throughput screening of miniature pig kidney N-glycan-binding proteins by using a mass-spectrometric approach. Without any derivatization steps, the purified pig kidney N-glycans were directly immobilized on to HBA-functionalized beads and subsequently used to identify GBPs from human serum. This screening method showed remarkable performance for identifying potential GBPs closely involved in pig-to-human xenograft rejection mediated by human serum, including antibodies, cytokines, complement components, siglec, and CD antigens. Thus, these results demonstrate that the GBP screening method was firmly established by one-step immobilization of the N-glycans on to microsphere and highly sensitive mass-spectrometric analysis. © 2008 Elsevier Ltd. All rights reserved.
  • Authors

    Digital Object Identifier (doi)

    Author List

  • Kim YG; Shin DS; Yang YH; Gil GC; Park CG; Mimura Y; Cooper DKC; Rudd PM; Dwek RA; Lee YS
  • Start Page

  • 215
  • End Page

  • 223
  • Volume

  • 15
  • Issue

  • 3