title: Comparative N-linked glycan analysis of wild-type and α1,3-galactosyltransferase gene knock-out pig fibroblasts using mass spectrometry approaches.
abstract: Carbohydrate antigens expressed on pig cells are considered to be major barriers in pig-to-human xenotransplantation. Even after α1,3-galactosyltransferase gene knock-out (GalT-KO) pigs are generated, potential non-Gal antigens are still existed. However, to the best of our knowledge there is no extensive study analyzing N-glycans expressed on the GalT-KO pig tissues or cells. Here, we identified and quantified totally 47 N-glycans from wild-type (WT) and GalT-KO pig fibroblasts using mass spectrometry. First, our results confirmed the absence of galactose-alpha-1,3-galactose (α-Gal) residue in the GalT-KO pig cells. Interestingly, we showed that the level of overall fucosylated N-glycans from GalT-KO pig fibroblasts is much higher than from WT pig fibroblasts. Moreover, the relative quantity of the N-glycolylneuraminic acid (NeuGc) antigen is slightly higher in the GalT-KO pigs. Thus, this study will contribute to a better understanding of cellular glycan alterations on GalT-KO pigs for successful xenotransplantation.
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Korea.
authors.forename: Hae-Min
authors.initials: HM
authors.lastname: Park
authors.affiliation: Department of Chemical Engineering, Soongsil University, Seoul 156-743, Korea.
authors.forename: Yoon-Woo
authors.initials: YW
authors.lastname: Kim
authors.affiliation: Department of Chemical Engineering, Soongsil University, Seoul 156-743, Korea.
authors.forename: Kyoung-Jin
authors.initials: KJ
authors.lastname: Kim
authors.affiliation: Department of Nanobiomedical Science and BK21+ NBM Global Research Center for Regenerative Medicine, Dankook University, Cheonan 330-714, Korea.
authors.forename: Young June
authors.initials: YJ
authors.lastname: Kim
authors.affiliation: Department of Microbial Engineering, College of Engineering, Konkuk University, Seoul 143-701, Korea.
authors.forename: Yung-Hun
authors.initials: YH
authors.lastname: Yang
authors.affiliation: Division of Mass Spectrometry Research, Korea Basic Science Institute, Ochang 363-883, Korea.
authors.forename: Jang Mi
authors.initials: JM
authors.lastname: Jin
authors.affiliation: Division of Mass Spectrometry Research, Korea Basic Science Institute, Ochang 363-883, Korea ; Department of Bio-Analytical Science, University of Science and Technology, Daejeon 305-764, Korea.
authors.forename: Young Hwan
authors.initials: YH
authors.lastname: Kim
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, Seoul 151-742, Korea.
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
authors.affiliation: Department of Nanobiomedical Science and BK21+ NBM Global Research Center for Regenerative Medicine, Dankook University, Cheonan 330-714, Korea.
authors.forename: Hosup
authors.initials: H
authors.lastname: Shim
authors.affiliation: Department of Chemical Engineering, Soongsil University, Seoul 156-743, Korea.
authors.forename: Yun-Gon
authors.initials: YG
authors.lastname: Kim
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology* / Q000276:immunology; D005700:Galactosyltransferases / Q000235:genetics* / Q000378:metabolism; D055786:Gene Knockout Techniques; D013058:Mass Spectrometry; D009438:Neuraminic Acids / Q000378:metabolism*; D011134:Polysaccharides / Q000302:isolation & purification*; D013552:Swine
chemical_list: D009438:Neuraminic Acids; D011134:Polysaccharides; C032592:N-glycolylneuraminic acid; D005700:Galactosyltransferases
keywords: GalT-KO pig fibroblast; N-glycan; N-glycolylneuraminic acid (NeuGc); mass spectrometry (MS)