Cx4Mab-1: A Novel Anti-Mouse CXCR4 Monoclonal Antibody for Flow Cytometry.

Q3 Medicine
Tsunenori Ouchida, Hiroyuki Suzuki, Tomohiro Tanaka, Mika K Kaneko, Yukinari Kato
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Abstract

The CXC chemokine receptor 4 (CXCR4, CD184) is a member of the G protein-coupled receptor family that is expressed in most leukocytes. Overexpression of CXCR4 is associated with poor prognosis in not only hematopoietic malignancy but also solid tumors. Because CXCR4 is an attractive target for tumor therapy, reliable preclinical murine models using anti-CXCR4 monoclonal antibodies (mAbs) have been warranted. This study established a novel anti-mouse CXCR4 (mCXCR4) mAb using the Cell-Based Immunization and Screening method. Flow cytometric analysis showed that an anti-mCXCR4 mAb, Cx4Mab-1 (rat IgG2a, kappa), recognized mCXCR4-overexpressed Chinese hamster ovary-K1 (CHO/mCXCR4) cells and endogenously mCXCR4-expressing mouse myeloma P3X63Ag8U.1 (P3U1) cells. Furthermore, Cx4Mab-1 did not recognize mCXCR4-knockout P3U1 cells. The dissociation constants of Cx4Mab-1 for CHO/mCXCR4 and P3U1 were determined as 6.4 × 10-9 M and 2.3 × 10-9 M, respectively, indicating that Cx4Mab-1 possesses a high affinity to both endogenous and exogenous mCXCR4-expressing cells. These results indicate that Cx4Mab-1 could be a useful tool for preclinical mouse models.

Cx4Mab-1:用于流式细胞仪的新型抗小鼠 CXCR4 单克隆抗体。
CXC 趋化因子受体 4(CXCR4,CD184)是 G 蛋白偶联受体家族的成员,在大多数白细胞中都有表达。CXCR4 的过表达不仅与造血恶性肿瘤的预后不良有关,也与实体瘤的预后不良有关。由于 CXCR4 是一个有吸引力的肿瘤治疗靶点,因此需要使用抗 CXCR4 单克隆抗体(mAbs)建立可靠的临床前小鼠模型。本研究利用细胞免疫和筛选方法建立了一种新型抗小鼠 CXCR4(mCXCR4)mAb。流式细胞分析表明,抗 mCXCR4 mAb Cx4Mab-1(大鼠 IgG2a,kappa)能识别过表达 mCXCR4 的中国仓鼠卵巢-K1(CHO/mCXCR4)细胞和内源性表达 mCXCR4 的小鼠骨髓瘤 P3X63Ag8U.1 (P3U1)细胞。此外,Cx4Mab-1 无法识别 mCXCR4 基因敲除的 P3U1 细胞。经测定,Cx4Mab-1 对 CHO/mCXCR4 和 P3U1 的解离常数分别为 6.4 × 10-9 M 和 2.3 × 10-9 M,这表明 Cx4Mab-1 对内源性和外源性表达 mCXCR4 的细胞都具有很高的亲和力。这些结果表明,Cx4Mab-1 可作为临床前小鼠模型的有用工具。
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
CiteScore
4.80
自引率
0.00%
发文量
49
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