title: High-throughput screening of glycan-binding proteins using miniature pig kidney N-glycan-immobilized beads.
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.
authors: Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, Seoul 151-742, Korea.
authors: Yun-Gon
authors: YG
authors: Kim
authors: Dong-Sik
authors: DS
authors: Shin
authors: Yung-Hun
authors: YH
authors: Yang
authors: Geun-Cheol
authors: GC
authors: Gil
authors: Chung-Gyu
authors: CG
authors: Park
authors: Yusuke
authors: Y
authors: Mimura
authors: David K C
authors: DK
authors: Cooper
authors: Pauline M
authors: PM
authors: Rudd
authors: Raymond A
authors: RA
authors: Dwek
authors: Yoon-Sik
authors: YS
authors: Lee
authors: Byung-Gee
authors: BG
authors: Kim
mesh_terms: D000818:Animals; D001565:Benzoates / Q000378:metabolism; D006084:Graft Rejection; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D013058:Mass Spectrometry; D008863:Microspheres; D011134:Polysaccharides / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013379:Substrate Specificity; D013552:Swine; D013556:Swine, Miniature; D014183:Transplantation, Heterologous / Q000276:immunology
chemical_list: D001565:Benzoates; D011134:Polysaccharides; D011506:Proteins; C053198:4-hydrazinobenzoic acid