Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
New Industry Creation Hatchery Center, Tohoku University, Sendai, Japan.
Monoclon Antib Immunodiagn Immunother. 2021 Jun;40(3):101-106. doi: 10.1089/mab.2021.0007.
CC chemokine receptor 9 (CCR9) belongs to the beta chemokine receptor family and is mainly distributed on the surface of immature T lymphocytes and enterocytes. This receptor is highly expressed in rheumatoid arthritis, colitis, type 2 diabetes, and various tumors. Therefore, more sensitive monoclonal antibodies (mAbs) need to be developed to predict the prognosis of many high CCR9 expression diseases. Because CCR9 is a structurally unstable G protein-coupled receptor, it has been difficult to develop anti-CCR9 mAbs using the traditional method. This study developed anti-human CCR9 (hCCR9) mAbs for flow cytometry using a Cell-Based Immunization and Screening (CBIS) method. Two mice were immunized with hCCR9-overexpressed Chinese hamster ovary (CHO)-K1 cells (CHO/hCCR9), and hybridomas showing strong signals from CHO/hCCR9 and no signals from CHO-K1 cells were selected by flow cytometry. We established an anti-hCCR9 mAb, CMab-1 (IgG, kappa), which detected hCCR9 in MOLT-4 leukemia T lymphoblast cells and CHO/hCCR9 cells by flow cytometry. Our study showed that an anti-hCCR9 mAb was developed more rapidly by the CBIS method than the previous method.
CC 趋化因子受体 9（CCR9）属于β趋化因子受体家族，主要分布于未成熟 T 淋巴细胞和肠上皮细胞表面。该受体在类风湿关节炎、结肠炎、2 型糖尿病和各种肿瘤中高度表达。因此，需要开发更敏感的单克隆抗体（mAbs）来预测许多高 CCR9 表达疾病的预后。由于 CCR9 是一种结构不稳定的 G 蛋白偶联受体，因此使用传统方法开发抗 CCR9 mAbs 一直具有挑战性。本研究使用基于细胞的免疫接种和筛选（CBIS）方法开发了用于流式细胞术的抗人 CCR9（hCCR9）mAbs。用 hCCR9 过表达的中国仓鼠卵巢（CHO）-K1 细胞（CHO/hCCR9）免疫了两只小鼠，通过流式细胞术选择对 CHO/hCCR9 显示强信号而对 CHO-K1 细胞无信号的杂交瘤。我们建立了抗 hCCR9 mAb CMab-1（IgG，kappa），该 mAb 通过流式细胞术检测 MOLT-4 白血病 T 淋巴母细胞和 CHO/hCCR9 细胞中的 hCCR9。我们的研究表明，与之前的方法相比，CBIS 方法可以更快地开发出抗 hCCR9 mAb。
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