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Comparative N-Linked Glycan Analysis of Wild-Type and α1,3-Galactosyltransferase Gene Knock-Out Pig Fibroblasts Using Mass Spectrometry Approaches

  • Park, Hae-Min (School of Chemical and Biological Engineering, Seoul National University) ;
  • Kim, Yoon-Woo (Department of Chemical Engineering, Soongsil University) ;
  • Kim, Kyoung-Jin (Department of Chemical Engineering, Soongsil University) ;
  • Kim, Young June (Department of Nanobiomedical Science and BK21+ NBM Global Research Center for Regenerative Medicine, Dankook University) ;
  • Yang, Yung-Hun (Department of Microbial Engineering, College of Engineering, Konkuk University) ;
  • Jin, Jang Mi (Division of Mass Spectrometry Research, Korea Basic Science Institute) ;
  • Kim, Young Hwan (Division of Mass Spectrometry Research, Korea Basic Science Institute) ;
  • Kim, Byung-Gee (School of Chemical and Biological Engineering, Seoul National University) ;
  • Shim, Hosup (Department of Nanobiomedical Science and BK21+ NBM Global Research Center for Regenerative Medicine, Dankook University) ;
  • Kim, Yun-Gon (Department of Chemical Engineering, Soongsil University)
  • Received : 2014.09.02
  • Accepted : 2014.10.21
  • Published : 2015.01.31

Abstract

Carbohydrate antigens expressed on pig cells are considered to be major barriers in pig-to-human xenotransplantation. Even after ${\alpha}1,3$-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 (${\alpha}$-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.

Keywords

GalT-KO pig fibroblast;mass spectrometry (MS);N-glycan;N-glycolylneuraminic acid (NeuGc)

Acknowledgement

Supported by : National Research Foundation of Korea (NRF)

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