S-Space College of Medicine/School of Medicine (의과대학/대학원) Microbiology (미생물학전공) Journal Papers (저널논문_미생물학전공)
High-throughput screening of glycan-binding proteins using miniature pig kidney N-glycan-immobilized beads
- Kim, Yun-Gon; Shin, Dong-Sik; Yang, Yung-Hun; Gil, Geun-Cheol; Park, Chung-Gyu; Mimura, Yusuke; Cooper, David K C; Rudd, Pauline M; Dwek, Raymond A; Lee, Yoon-Sik; Kim, Byung-Gee
- Issue Date
- Chem Biol. 2008 Mar;15(3):215-23.
- Animals; Benzoic Acids/metabolism; Graft Rejection; Humans; Kidney/*metabolism; Mass Spectrometry; *Microspheres; Polysaccharides/*metabolism; Protein Binding; Proteins/*metabolism; Substrate Specificity; Swine; *Swine, Miniature; Transplantation, Heterologous/immunology
- 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.
- 1074-5521 (Print)
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