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High-throughput screening of glycan-binding proteins using miniature pig kidney N-glycan-immobilized beads

Cited 12 time in Web of Science Cited 14 time in Scopus
Authors
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
2008-03-22
Publisher
Elsevier
Citation
Chem Biol. 2008 Mar;15(3):215-23.
Keywords
AnimalsBenzoic Acids/metabolismGraft RejectionHumansKidney/*metabolismMass Spectrometry*MicrospheresPolysaccharides/*metabolismProtein BindingProteins/*metabolismSubstrate SpecificitySwine*Swine, MiniatureTransplantation, Heterologous/immunology
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.
ISSN
1074-5521 (Print)
Language
English
URI
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Citation&list_uids=18355721

http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6VRP-4S3PX1T-8-2&_cdi=6240&_user=168665&_orig=search&_coverDate=03%2F21%2F2008&_sk=999849996&view=c&wchp=dGLbVlz-zSkzV&md5=171bc39c8174db3bc82504d61ac4b07d&ie=/sdarticle.pdf

http://hdl.handle.net/10371/45519
DOI
https://doi.org/10.1016/j.chembiol.2008.02.009
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College of Medicine/School of Medicine (의과대학/대학원)Microbiology (미생물학전공)Journal Papers (저널논문_미생물학전공)
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