抗鼠 CCR3 单克隆抗体（CMab-6 和 CMab-7）表位作图。
Epitope Mapping of Anti-Mouse CCR3 Monoclonal Antibodies (CMab-6 and CMab-7).
机构信息
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Department of Molecular Pharmacology, and Tohoku University Graduate School of Medicine, Sendai, Japan.
出版信息
Monoclon Antib Immunodiagn Immunother. 2023 Apr;42(2):68-72. doi: 10.1089/mab.2022.0034. Epub 2023 Apr 18.
One of G protein-coupled receptors, CC chemokine receptor 3 (CCR3), is expressed in eosinophils, basophils, a subset of Th2 lymphocytes, mast cells, and airway epithelial cells. CCR3 levels in the serum of colorectal cancer patients are significantly higher than in control groups. Moreover, CCR3 is essential for recruiting eosinophils into the lung. Therefore, CCR3 is considered both a therapeutic target for colorectal cancer and allergic diseases. Previously, we established anti-mouse CCR3 (mCCR3) monoclonal antibodies (mAbs), CMab-6 (rat IgG, kappa) and CMab-7 (rat IgG, kappa), by immunizing a rat with an N-terminal peptide of mCCR3. These mAbs can be used in flow cytometry and enzyme-linked immunosorbent assays. In this study, we performed the epitope mapping of CMab-6 and CMab-7 using alanine scanning. The reactivity between these mAbs and point mutants of mCCR3 were analyzed using flow cytometry. The results indicated that Phe3, Asn4, Thr5, Asp6, Glu7, Lys9, Thr10, and Glu13 of mCCR3 are essential for CMab-6 binding, whereas Phe15 and Glu16 are essential for CMab-7 binding.
G 蛋白偶联受体之一，CC 趋化因子受体 3（CCR3），在嗜酸性粒细胞、嗜碱性粒细胞、Th2 淋巴细胞亚群、肥大细胞和气道上皮细胞中表达。结直肠癌患者血清中的 CCR3 水平明显高于对照组。此外，CCR3 对于将嗜酸性粒细胞招募到肺部至关重要。因此，CCR3 被认为既是结直肠癌的治疗靶点，也是过敏疾病的治疗靶点。此前，我们通过用 mCCR3 的 N 端肽免疫大鼠，建立了抗小鼠 CCR3（mCCR3）单克隆抗体（mAbs）CMab-6（大鼠 IgG，kappa）和 CMab-7（大鼠 IgG，kappa）。这些 mAbs 可用于流式细胞术和酶联免疫吸附测定。在这项研究中，我们使用丙氨酸扫描法对 CMab-6 和 CMab-7 进行了表位作图。通过流式细胞术分析了这些 mAbs 与 mCCR3 点突变体之间的反应性。结果表明，mCCR3 的 Phe3、Asn4、Thr5、Asp6、Glu7、Lys9、Thr10 和 Glu13 对于 CMab-6 的结合是必需的，而 Phe15 和 Glu16 对于 CMab-7 的结合是必需的。
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