A Rat Anti-Mouse CD39 Monoclonal Antibody for Flow Cytometry.

Q3 Medicine Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2023-12-01 Epub Date: 2023-12-21 DOI:10.1089/mab.2023.0018

Hiroyuki Suzuki, Tomohiro Tanaka, Yuma Kudo, Mayuki Tawara, Aoi Hirayama, Mika K Kaneko, Yukinari Kato

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Abstract

By converting extracellular adenosine triphosphate to adenosine, CD39 is involved in adenosine metabolism. The extracellular adenosine plays a critical role in the immune suppression of the tumor microenvironment. Therefore, the inhibition of CD39 activity by monoclonal antibodies (mAbs) is thought to be one of the important strategies for tumor therapy. In this study, we developed novel mAbs for mouse CD39 (mCD39) using the Cell-Based Immunization and Screening (CBIS) method. One of the established anti-mCD39 mAbs, C₃₉Mab-2 (rat IgG_2a, lambda), reacted with mCD39-overexpressed Chinese hamster ovary-K1 (CHO/mCD39) and an endogenously mCD39-expressed cell line (SN36) by flow cytometry. The kinetic analysis using flow cytometry indicated that the dissociation constant (K_D) values of C₃₉Mab-2 for CHO/mCD39 and SN36 were 5.5 × 10^-9 M and 4.9 × 10^-9 M, respectively. These results indicated that C₃₉Mab-2 is useful for the detection of mCD39 in flow cytometry.

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用于流式细胞仪的大鼠抗小鼠 CD39 单克隆抗体。

通过将细胞外的三磷酸腺苷转化为腺苷，CD39 参与了腺苷代谢。细胞外腺苷在肿瘤微环境的免疫抑制中起着至关重要的作用。因此，通过单克隆抗体（mAbs）抑制 CD39 的活性被认为是肿瘤治疗的重要策略之一。在这项研究中，我们利用细胞免疫和筛选（CBIS）方法开发了新型小鼠 CD39（mCD39）mAbs。已建立的抗 mCD39 mAbs 之一 C39Mab-2（大鼠 IgG2a，λ）通过流式细胞术与过表达 mCD39 的中国仓鼠卵巢-K1（CHO/mCD39）和内源性 mCD39 表达的细胞系（SN36）反应。流式细胞仪的动力学分析表明，C39Mab-2 对 CHO/mCD39 和 SN36 的解离常数（KD）分别为 5.5 × 10-9 M 和 4.9 × 10-9 M。这些结果表明，C39Mab-2 在流式细胞仪中可用于检测 mCD39。

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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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