Development of Anti-Human CC Chemokine Receptor 9 Monoclonal Antibodies for Flow Cytometry
Development of Anti-Human CC Chemokine Receptor 9 Monoclonal Antibodies for Flow Cytometry
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DOI：
10.1089/mab.2021.0007
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发表时间：
2021-06-01
影响因子：
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通讯作者：
Kato, Yukinari
中科院分区：
文献类型：
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作者：
Nanamiya, Ren;Takei, Junko;Kato, Yukinari
CC趋化因子受体9（CCR 9）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。这种受体在类风湿性关节炎、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（mAb）来预测许多CCR 9高表达疾病的预后。由于CCR 9是一种结构不稳定的G蛋白偶联受体，使用传统方法很难开发抗CCR 9的mAb。本研究使用基于细胞的免疫和筛选（CBIS）方法开发用于流式细胞术的抗人CCR 9（hCCR 9）mAb。用hCCR 9过表达的中国仓鼠卵巢（CHO）-K1细胞（CHO/hCCR 9）免疫两只小鼠，并通过流式细胞术选择显示来自CHO/hCCR 9的强信号和不显示来自CHO-K1细胞的信号的杂交瘤。我们建立了抗hCCR 9 mAb，C（9）Mab-1（IgG（1），kappa），通过流式细胞术检测MOLT-4白血病T淋巴母细胞和CHO/hCCR 9细胞中的hCCR 9。我们的研究表明，抗hCCR 9单克隆抗体的CBIS方法比以前的方法更快地开发。
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, C(9)Mab-1 (IgG(1), 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.