Cx1Mab-1: A Novel Anti-mouse CXCR1 Monoclonal Antibody for Flow Cytometry.

Q3 Medicine Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2024-04-01 DOI:10.1089/mab.2023.0031

Guanjie Li, Tomohiro Tanaka, Tsunenori Ouchida, M. Kaneko, Hiroyuki Suzuki, Y. Kato

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Abstract

The C-X-C motif chemokine receptor-1 (CXCR1) is a rhodopsin-like G-protein-coupled receptor, expressed on the cell surface of immune cells and tumors. CXCR1 interacts with some C-X-C chemokines, such as CXCL6, CXCL7, and CXCL8/interleukin-8, which are produced by various cells. Since CXCR1 is involved in several diseases including tumors and diabetes mellitus, drugs targeting CXCR1 have been developed. Therefore, the development of sensitive monoclonal antibodies (mAbs) for CXCR1 has been desired for the diagnosis and treatment. This study established a novel anti-mouse CXCR1 (mCXCR1) mAb, Cx1Mab-1 (rat IgG1, kappa), using the Cell-Based Immunization and Screening method. Cx1Mab-1 reacted with mCXCR1-overexpressed Chinese hamster ovary-K1 (CHO/mCXCR1) and mCXCR1-overexpressed LN229 glioblastoma (LN229/mCXCR1) in flow cytometry. Cx1Mab-1 demonstrated a high binding affinity for CHO/mCXCR1 and LN229/mCXCR1 with a dissociation constant of 2.6 × 10-9 M and 2.1 × 10-8 M, respectively. Furthermore, Cx1Mab-1 could detect mCXCR1 by Western blot analysis. These results indicated that Cx1Mab-1 is useful for detecting mCXCR1, and provides a possibility for targeting mCXCR1-expressing cells in vivo experiments.

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Cx1Mab-1：用于流式细胞仪的新型抗小鼠 CXCR1 单克隆抗体。

C-X-C motif趋化因子受体-1（CXCR1）是一种类似于红细胞蛋白的G蛋白偶联受体，表达于免疫细胞和肿瘤的细胞表面。CXCR1 与各种细胞产生的一些 C-X-C 趋化因子（如 CXCL6、CXCL7 和 CXCL8/白细胞介素-8）相互作用。由于 CXCR1 与包括肿瘤和糖尿病在内的多种疾病有关，针对 CXCR1 的药物已被开发出来。因此，人们一直希望开发出敏感的 CXCR1 单克隆抗体（mAbs），用于诊断和治疗。本研究利用细胞免疫和筛选方法建立了一种新型抗小鼠 CXCR1（mCXCR1）mAb--Cx1Mab-1（大鼠 IgG1，kappa）。在流式细胞术中，Cx1Mab-1 与表达 mCXCR1 的中国仓鼠卵巢-K1（CHO/mCXCR1）和表达 mCXCR1 的 LN229 胶质母细胞瘤（LN229/mCXCR1）发生了反应。Cx1Mab-1 与 CHO/mCXCR1 和 LN229/mCXCR1 的结合亲和力很高，解离常数分别为 2.6 × 10-9 M 和 2.1 × 10-8 M。此外，Cx1Mab-1 还能通过 Western 印迹分析检测到 mCXCR1。这些结果表明，Cx1Mab-1 可用于检测 mCXCR1，并为体内实验中靶向表达 mCXCR1 的细胞提供了可能。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Medicine-Immunology and Allergy

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4.80

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