Specific detection of Neu5Gc in animal tissues by immunohistochemistry

Anu Paul, Shirley Bachar Abramovitch, Vered Padler-Karavani

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

One of the major obstacles in xenotransplantation is tissue expression of the non-human mammalian carbohydrate N-glycolylneuraminic acid (Neu5Gc). This 9-carbon backbone acidic sugar is the hydroxylated form of N-acetylneuraminic acid (Neu5Ac), and both constitute the two most common sialic acid types in mammals. Loss of CMP-Neu5Ac hydroxylase encoding gene in humans dictates the immunogenic nature of Neu5Gc-containing xenografts. Here we describe an immunohistochemistry method for the detection of Neu5Gc in mammalian-derived tissues using affinity-purified chicken anti-Neu5Gc IgY. Specificity is further demonstrated by competitive inhibition with free Neu5Gc or Neu5Gc-glycoproteins, but not with Neu5Ac or Neu5Ac-glycoproteins. This method can be used to evaluate potential Neu5Gc-immunogenicity of xenografts.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages59-72
Number of pages14
DOIs
StatePublished - 2020

Publication series

NameMethods in Molecular Biology
Volume2110
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Anti-carbohydrate antibodies
  • Immunohistochemistry
  • N-acetylneuraminic acid (Neu5Ac)
  • N-glycolylneuraminic acid (Neu5Gc)
  • Sialic acid
  • Xenotransplantation

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