通过 N 端肽免疫建立新型抗小鼠 CCR3 单克隆抗体（CMab-6 和 CMab-7）。
Establishment of Novel Anti-Mouse CCR3 Monoclonal Antibodies (CMab-6 and CMab-7) by N-terminal Peptide Immunization.
机构信息
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, Sendai, Japan.
出版信息
Monoclon Antib Immunodiagn Immunother. 2022 Apr;41(2):94-100. doi: 10.1089/mab.2021.0065.
The CC chemokine receptor 3 (CCR3) is a member of the G protein-coupled receptor family that is highly expressed in eosinophils and basophils. CCR3 has been proposed as a therapeutic target for human immunodeficiency virus and allergy diagnosis. Therefore, in this study, we developed specific and sensitive monoclonal antibodies (mAbs) for mouse CCR3 (mCCR3), which are useful for flow cytometry by peptide immunization. The established anti-mCCR3 mAbs, CMab-6 (rat IgG, kappa) and CMab-7 (rat IgG, kappa), reacted with mCCR3-overexpressed Chinese hamster ovary-K1 (CHO/mCCR3), in addition to mCCR3-endogenously expressed cell lines, such as P388 (mouse lymphoid neoplasma) and J774-1 (mouse macrophage-like) through flow cytometry. Kinetic analyses using flow cytometry indicated that the dissociation constants (s) of CMab-6 for CHO/mCCR3, P388, and J774-1 cells were 8.7 × 10 M, 1.4 × 10 M, and 1.7 × 10 M, respectively, whereas the s of CMab-7 for these cell lines were 3.7 × 10 M, 5.1 × 10 M, and 3.1 × 10 M, respectively. Results also indicated that CMab-6 and CMab-7 are useful for detecting cells expressing CCR3 through flow cytometry, thereby making them potentially beneficial for treating CCR3-expressing cells.
CC 趋化因子受体 3（CCR3）是 G 蛋白偶联受体家族的成员，在嗜酸性粒细胞和嗜碱性粒细胞中高度表达。CCR3 已被提议作为人类免疫缺陷病毒和过敏诊断的治疗靶点。因此，在这项研究中，我们通过肽免疫开发了针对小鼠 CCR3（mCCR3）的特异性和敏感的单克隆抗体（mAbs），这些 mAbs 可用于流式细胞术。建立的抗 mCCR3 mAbs，CMab-6（大鼠 IgG，kappa）和 CMab-7（大鼠 IgG，kappa），通过流式细胞术与 mCCR3 过表达的中国仓鼠卵巢-K1（CHO/mCCR3）以及 mCCR3 内源性表达的细胞系（如 P388（小鼠淋巴样肿瘤）和 J774-1（小鼠巨噬样））反应。使用流式细胞术进行的动力学分析表明，CMab-6 对 CHO/mCCR3、P388 和 J774-1 细胞的解离常数（s）分别为 8.7×10-9 M、1.4×10-9 M 和 1.7×10-9 M，而 CMab-7 对这些细胞系的 s 分别为 3.7×10-9 M、5.1×10-9 M 和 3.1×10-9 M。结果还表明，CMab-6 和 CMab-7 可用于通过流式细胞术检测表达 CCR3 的细胞，从而使其对治疗表达 CCR3 的细胞可能有益。
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