A novel anti-mouse CCR7 monoclonal antibody, C7Mab-7, demonstrates high sensitivity in flow cytometry, western blot, and immunohistochemistry

IF 2.2 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Biochemistry and Biophysics Reports Pub Date : 2025-03-01 Epub Date: 2025-02-11 DOI:10.1016/j.bbrep.2025.101948

Hiroyuki Satofuka, Hiroyuki Suzuki, Tomohiro Tanaka, Rena Ubukata, Miu Hirose, Haruto Yamamoto, Yu Kaneko, Shiori Fujisawa, Guanjie Li, Mika K. Kaneko, Yukinari Kato

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Abstract

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, C₇Mab-7 (rat IgG₁, kappa) using the Cell-Based Immunization and Screening (CBIS) method. C₇Mab-7 demonstrated high sensitivity in flow cytometry. The dissociation constant (K_D) value of C₇Mab-7 was determined to be 2.5 × 10⁻⁹ M for mCCR7-overexpressed Chinese hamster ovary-K1 (CHO/mCCR7) cells. Furthermore, C₇Mab-7 detected mCCR7 with high sensitivity in western blot and immunohistochemistry. C₇Mab-7, developed by the CBIS method, accelerates the development of CCR7-targeted antibody therapies and cancer diagnostics.

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一种新的抗小鼠CCR7单克隆抗体C7Mab-7在流式细胞术、western blot和免疫组织化学中显示出高灵敏度

C-C趋化因子受体7型（CCR7）是G蛋白偶联受体家族的成员，是免疫细胞的淋巴结归巢受体。在配体结合后，CCR7促进免疫细胞向次级淋巴器官的迁移。在癌症中，CCR7已被发现是淋巴结转移的关键分子。因此，抗ccr7单克隆抗体（mab）已被开发为癌症治疗药物。本研究采用细胞免疫筛选（CBIS）方法建立抗小鼠CCR7 （mCCR7）单抗、C7Mab-7（大鼠IgG1、kappa）单抗。C7Mab-7在流式细胞术中表现出高灵敏度。C7Mab-7对mCCR7过表达的中国仓鼠卵巢k1 （CHO/mCCR7）细胞的解离常数（KD）为2.5 × 10⁻(9)M。此外，C7Mab-7在western blot和免疫组织化学中检测mCCR7具有较高的敏感性。C7Mab-7由CBIS方法开发，加速了ccr7靶向抗体疗法和癌症诊断的发展。

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来源期刊

Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

191

审稿时长

59 days

期刊介绍： Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.