TY - CHAP
T1 - Specific detection of Neu5Gc in animal tissues by immunohistochemistry
AU - Paul, Anu
AU - Bachar Abramovitch, Shirley
AU - Padler-Karavani, Vered
N1 - Publisher Copyright:
© Springer Science+Business Media, LLC, part of Springer Nature 2020.
PY - 2020
Y1 - 2020
N2 - 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.
AB - 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.
KW - Anti-carbohydrate antibodies
KW - Immunohistochemistry
KW - N-acetylneuraminic acid (Neu5Ac)
KW - N-glycolylneuraminic acid (Neu5Gc)
KW - Sialic acid
KW - Xenotransplantation
UR - http://www.scopus.com/inward/record.url?scp=85078710805&partnerID=8YFLogxK
U2 - 10.1007/978-1-0716-0255-3_4
DO - 10.1007/978-1-0716-0255-3_4
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C2 - 32002901
AN - SCOPUS:85078710805
T3 - Methods in Molecular Biology
SP - 59
EP - 72
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -