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Generation of monoclonal antibodies against the Galβ1-4Gal epitope: a key tool in studies of species-specific glycans expressed in fish, amphibians, and birds.
MedLine Citation:
PMID:  22948803     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Whereas the Galβ1-4Gal epitope is rarely found in mammalian glycans, it has been found in glycans of various species of non-mammalian vertebrates, such as fish, amphibians, and birds. Although glycans containing Galβ1-4Gal in these vertebrates were detected by precise structural analysis of the glycans using mass spectrometry and/or NMR spectrometry, there are no convenient methods to detect Galβ1-4Gal from various samples. To analyze systematically the distribution of Galβ1-4Gal in nature, we generated mouse monoclonal antibodies (mAbs) specific for Galβ1-4Gal using extracts of medaka eggs as an immunogen. Four mAbs (two IgMs and two IgG1s) were obtained by ELISA-based screening. The specificities of these mAbs were evaluated by frontal affinity chromatography using 142 kinds of 2-aminopyridine-derivatized oligosaccharides. While all mAbs interacted with (Galβ1-4Gal)-containing oligosaccharides at their non-reducing termini with dissociation constants (K(d)) ranging from 1.0x10(-5) to 2.8x10(-4) M, no apparent interaction was observed with any other glycans. The number of branches containing Galβ1-4Gal on N-glycans did not significantly affect K(d) of mAbs of IgG1-subclasses, but those of IgM-mAbs were decreased by about one order of magnitude, in increments of the number of branches present. Using the mAbs, we established that Galβ1-4Gal is also expressed on glycoproteins in various tissues from the African clawed frog. Immunohistocheminal staining of medaka sections revealed that Galβ1-4Gal epitopes were expressed in the endothelium, epithelium, and epidermis, which directly contact the external environment or invading organisms. Thus, these mAbs are useful for systematically investigating the species-specific expression of glycans, which may act as a barrier against infection.
Authors:
Noriko Suzuki; Daisuke Nawa; Hiroaki Tateno; Takako Yasuda; Shoji Oda; Hiroshi Mitani; Toshiyuki Nishimaki; Takafumi Katsumura; Hiroki Oota; Tsunehiko Hanihara; Atsunori Oga; Jun Hirabayashi; Kazuo Yamamoto
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-9-3
Journal Detail:
Title:  Glycobiology     Volume:  -     ISSN:  1460-2423     ISO Abbreviation:  Glycobiology     Publication Date:  2012 Sep 
Date Detail:
Created Date:  2012-9-5     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9104124     Medline TA:  Glycobiology     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Chiba, 277-8562, Japan.
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