Earticle

초록

영어

Glycan recognitionleading 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. Firstly, total N-glycans from membrane glycoproteins derived from specific pathogen-free miniature pig kidney were qualitatively and quantitatively identified by MALDI-TOF, negative ion ESI MS/MS and normal-phase HPLC (NP-HPLC) combined with exoglycosidase digestion. Over 100 N-glycans, including sialylated and neutral types, were identified. Without any derivatization steps, the characterizedpig 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.

저자

• Yun-Gon Kim [ Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University ]
• Dong-Sik Shin [ School of Chemical and Biological Engineering, Seoul National University, Seoul ]
• David J. Harvey [ Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom ]
• Hee-Jin Jeong [ School of Chemical and Biological Engineering, Seoul National University, Seoul ]
• Kyoung-Soon Jang [ Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University ]
• Yung-Hun Yang [ School of Chemical and Biological Engineering, Seoul National University ]
• Chung-Gyu Park [ Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul ]
• Pauline M. Rudd [ Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, United Kingdom, The Dublin-Oxford Glycobiology Laboratory, National Institute for Bioprocessing Research and Training, Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland ]
• Raymond A. Dwek [ The Dublin-Oxford Glycobiology Laboratory, National Institute for Bioprocessing Research and Training, Conway Institute, University College Dublin, Belfield, Dublin 4, Ireland ]
• Yoon-Sik Lee [ School of Chemical and Biological Engineering, Seoul National University, Seoul ]
• Byung-Gee Kim [ Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University, School of Chemical and Biological Engineering, Seoul National University, Seoul ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국당과학회 학술대회

• 간기

  연간

• 수록기간

  2006~2022

• 십진분류

  KDC 517 DDC 614