Abstract
Anti-carbohydrate monoclonal antibodies (mAbs) are very useful in the functional analysis of complex carbohydrates in vivo. However, such mAbs are difficult to generate, largely because a wide variety of complex carbohydrates is intrinsically expressed in mice and rats and because the antigenicities of glycans are generally poor. In this chapter, I describe an efficient method for generating anti-carbohydrate mAbs using glycan-synthesizing enzyme-knockout mice in which the glycan structures formed by the missing enzymes should be highly antigenic. As an application of this method, I describe the generation of anti-sulfated glycan mAbs using sulfotransferase-deficient mice and the immunohistochemical detection of sulfated glycans involved in lymphocyte homing in both humans and mice.
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Acknowledgments
I would like to thank Drs. Jotaro Hirakawa (University of Shizuoka, School of Pharmaceutical Sciences), Minoru Fukuda (Sanford-Burnham Medical Research Institute), and Motohiro Kobayashi (Shinshu University, School of Medicine) for collaboration. This work was supported in part by Grants-in-Aid for Scientific Research, Category (B) and Grants-in-Aid for Scientific Research on Priority Areas, Dynamics of Extracellular Environments, from the Ministry of Education, Culture, Sports, Science and Technology, Japan (21390023 and 20057022, respectively).
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Kawashima, H. (2013). Generation of Anti-sulfated Glycan Antibodies Using Sulfotransferase-Deficient Mice. In: Brockhausen, I. (eds) Glycosyltransferases. Methods in Molecular Biology, vol 1022. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-465-4_5
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DOI: https://doi.org/10.1007/978-1-62703-465-4_5
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