Satofuka Hiroyuki, Suzuki Hiroyuki, Tanaka Tomohiro, Ubukata Rena, Hirose Miu, Yamamoto Haruto, Kaneko Yu, Fujisawa Shiori, Li Guanjie, Kaneko Mika K, Kato Yukinari
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, Japan.
Biochem Biophys Rep. 2025 Feb 11;41:101948. doi: 10.1016/j.bbrep.2025.101948. eCollection 2025 Mar.
C-C chemokine receptor type 7 (CCR7) is a member of the G protein-coupled receptor family and functions as a lymph node-homing receptor for immune cells. Upon ligand binding, CCR7 promotes the migration of immune cells to secondary lymphoid organs. In cancers, CCR7 has been revealed as a critical molecule in lymph node metastasis. Consequently, anti-CCR7 monoclonal antibodies (mAbs) have been developed as cancer therapeutic agents. In this study, we established an anti-mouse CCR7 (mCCR7) mAb, CMab-7 (rat IgG, kappa) using the Cell-Based Immunization and Screening (CBIS) method. CMab-7 demonstrated high sensitivity in flow cytometry. The dissociation constant ( ) value of CMab-7 was determined to be 2.5 × 10⁻⁹ M for mCCR7-overexpressed Chinese hamster ovary-K1 (CHO/mCCR7) cells. Furthermore, CMab-7 detected mCCR7 with high sensitivity in western blot and immunohistochemistry. CMab-7, developed by the CBIS method, accelerates the development of CCR7-targeted antibody therapies and cancer diagnostics.
C-C趋化因子受体7（CCR7）是G蛋白偶联受体家族的成员，作为免疫细胞的淋巴结归巢受体发挥作用。在配体结合后，CCR7促进免疫细胞向次级淋巴器官迁移。在癌症中，CCR7已被揭示为淋巴结转移中的关键分子。因此，抗CCR7单克隆抗体（mAb）已被开发为癌症治疗药物。在本研究中，我们使用基于细胞的免疫和筛选（CBIS）方法建立了一种抗小鼠CCR7（mCCR7）单克隆抗体CMab-7（大鼠IgG，κ）。CMab-7在流式细胞术中表现出高灵敏度。对于过表达mCCR7的中国仓鼠卵巢-K1（CHO/mCCR7）细胞，CMab-7的解离常数（）值测定为2.5×10⁻⁹M。此外，CMab-7在蛋白质印迹和免疫组织化学中以高灵敏度检测到mCCR7。通过CBIS方法开发的CMab-7加速了CCR7靶向抗体疗法和癌症诊断的发展。
