Qualitative and quantitative comparison of N‐glycans between pig endothelial and islet cells by high‐performance liquid chromatography and mass spectrometry‐based strategy |
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Authors: | Yun‐Gon Kim Geun‐Cheol Gil Kyung‐Soon Jang Sukmook Lee Hyoung‐Il Kim Jung‐Sik Kim Junho Chung Chung‐Gyu Park David J Harvey Byung‐Gee Kim |
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Institution: | 1. Institute of Molecular Biology and Genetics, Seoul National University, Seoul, Korea;2. Institute of Bioengineering, Seoul National University, Seoul, Korea;3. School of Chemical and Biological Engineering in College of Engineering, Seoul National University, Seoul, Korea;4. Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul, Korea;5. Xenotransplantation Research Center and Transplantation Research Institute, Seoul National University College of Medicine, Seoul, Korea;6. Department of Microbiology and Immunology, Seoul National University College of Medicine, Seoul, Korea;7. TIMRC and Cancer Research Institutue, Seoul National University College of Medicine, Seoul, Korea;8. Oxford Glycobiology Institute, Department of Biochemistry, University of Oxford, Oxford, UK |
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Abstract: | N‐glycan structures released from miniature pig endothelial and islet cells were determined by matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI‐TOF), negative ion electrospray ionization (ESI) MS/MS and normal‐phase high performance liquid chromatography (NP‐HPLC) combined with exoglycosidase digestion. Totally, the identified structures were 181 N‐glycans including 129 sialylated and 18 α‐galactosylated glycans from pig endothelial cells and 80 N‐glycans including 41 sialylated and one α‐galactosylated glycans from pig islet cells. The quantity of the α‐galactosylated glycans from pig islet cells was certainly neglectable compared to pig endothelial cells. A number of NeuGc‐terminated N‐glycans (80 from pig endothelial cells and 13 from pig islet cells) are newly detected by our mass spectrometric strategies. The detailed structural information will be a matter of great interest in organ or cell xenotransplantation using α 1,3‐galactosyltransferase gene‐knockout (GalT‐KO) pig. Copyright © 2009 John Wiley & Sons, Ltd. |
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Keywords: | xenotransplantation mass spectrometry α ‐galactose N‐glycolyl‐neuraminic acid pig endothelial cell pig islet cells |
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