Qualitative and quantitative comparison of N-glycans between pig endothelial and islet cells by high-performance liquid chromatography and mass spectrometry-based strategy

• Published in: Journal of mass spectrometry. 2009-07, Vol.44 (7), p.1087-1104
• Main Authors: Kim, Yun-Gon, Gil, Geun-Cheol, Jang, Kyung-Soon, Lee, Sukmook, Kim, Hyoung-Il, Kim, Jung-Sik, Chung, Junho, Park, Chung-Gyu, Harvey, David J, Kim, Byung-Gee
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Carbohydrates; Cell Line; Chromatography, High Pressure Liquid - methods; Endothelial cells; Endothelial Cells - chemistry; Endothelial Cells - metabolism; Fundamental and applied biological sciences. Psychology; Glycan; Glycomics - methods; Glycoside Hydrolases - metabolism; Investigative techniques, diagnostic techniques (general aspects); Ionization; Islets of Langerhans - chemistry; Islets of Langerhans - metabolism; Lasers; mass spectrometry; Medical sciences; Miniature; Miscellaneous; Miscellaneous. Technology; N-glycolyl-neuraminic acid; Other biological molecules; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; pig endothelial cell; pig islet cells; Polysaccharides - chemistry; Polysaccharides - metabolism; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Spectroscopy; Strategy; Swine; Xenotransplantation; α-galactose
• ISSN: 1076-5174; 1096-9888; 1096-9888
• DOI: 10.1002/jms.1587

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.