Kawashima Hiroto
Laboratory of Microbiology and Immunology, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan.
Methods Mol Biol. 2013;1022:51-60. doi: 10.1007/978-1-62703-465-4_5.
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.
抗碳水化合物单克隆抗体(mAb)在体内复杂碳水化合物的功能分析中非常有用。然而,此类单克隆抗体难以产生,主要原因是小鼠和大鼠体内天然表达多种复杂碳水化合物,且聚糖的抗原性通常较差。在本章中,我描述了一种利用聚糖合成酶基因敲除小鼠产生抗碳水化合物单克隆抗体的有效方法,在这种小鼠中,缺失酶所形成的聚糖结构应具有高度抗原性。作为该方法的一个应用,我描述了利用硫酸转移酶缺陷小鼠产生抗硫酸化聚糖单克隆抗体,以及在人和小鼠中对参与淋巴细胞归巢的硫酸化聚糖进行免疫组织化学检测。