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.
Key words
- Sialic acid
- N-acetylneuraminic acid (Neu5Ac)
- N-glycolylneuraminic acid (Neu5Gc)
- Anti-carbohydrate antibodies
- Xenotransplantation
- Immunohistochemistry
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Acknowledgment
This work was supported by the 7th Framework Program FP7-Health-2013-INNOVATION-1-603049 of the European Commission (to V.P-K).
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Paul, A., Bachar Abramovitch, S., Padler-Karavani, V. (2020). Specific Detection of Neu5Gc in Animal Tissues by Immunohistochemistry. In: Costa, C. (eds) Xenotransplantation. Methods in Molecular Biology, vol 2110. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0255-3_4
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DOI: https://doi.org/10.1007/978-1-0716-0255-3_4
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