Comparative N-linked glycan analysis of wild-type and α1,3-galactosyltransferase gene knock-out pig fibroblasts using mass spectrometry approaches

Mol Cells. 2015 Jan 31;38(1):65-74. doi: 10.14348/molcells.2015.2240. Epub 2014 Dec 16.

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.

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

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane / metabolism
  • Cells, Cultured
  • Fibroblasts / cytology
  • Fibroblasts / enzymology*
  • Fibroblasts / immunology
  • Galactosyltransferases / genetics*
  • Galactosyltransferases / metabolism
  • Gene Knockout Techniques
  • Mass Spectrometry
  • Neuraminic Acids / metabolism*
  • Polysaccharides / isolation & purification*
  • Swine

Substances

  • Neuraminic Acids
  • Polysaccharides
  • N-glycolylneuraminic acid
  • Galactosyltransferases