Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Bunkyo-ku, Japan.
Monoclon Antib Immunodiagn Immunother. 2021 Jun;40(3):124-133. doi: 10.1089/mab.2021.0011. Epub 2021 May 26.
The development of protein-specific antibodies is essential for understanding a wide variety of biological phenomena. Parasitic and viral infections and cancers are known to occur within California sea lion () populations. However, sensitive and specific monoclonal antibodies (mAbs) for the pathophysiological analysis of California sea lion tissues have not yet been developed. A type I transmembrane glycoprotein, podoplanin (PDPN), is a known diagnostic marker of lymphatic endothelial cells. We have previously developed several anti-PDPN mAbs in various mammalian species, with applications in flow cytometry, Western blotting, and immunohistochemistry. In this study, we established a novel mAb against California sea lion PDPN (seaPDPN), clone PMab-269 (mouse IgG, kappa), using a Cell-Based Immunization and Screening method. PMab-269 is specifically detected in seaPDPN-overexpressed Chinese hamster ovary (CHO)-K1 cells using flow cytometry and Western blotting. Moreover, PMab-269 clearly identified pulmonary type I alveolar cells, renal podocytes, and colon lymphatic endothelial cells in California sea lion tissues using immunohistochemistry. These findings demonstrate the usefulness of PMab-269 for the pathophysiological analysis of lung, kidney, and lymphatic tissues of the California sea lion.
蛋白特异性抗体的开发对于理解广泛的生物学现象至关重要。已知寄生虫和病毒感染以及癌症会在加利福尼亚海狮种群中发生。然而，用于加利福尼亚海狮组织病理生理学分析的敏感和特异性单克隆抗体（mAb）尚未开发。一种 I 型跨膜糖蛋白，足突蛋白（PDPN），是淋巴内皮细胞的已知诊断标志物。我们之前在各种哺乳动物物种中开发了几种抗 PDPN mAb，可用于流式细胞术、Western blot 和免疫组织化学。在这项研究中，我们使用基于细胞的免疫接种和筛选方法建立了一种针对加利福尼亚海狮 PDPN（seaPDPN）的新型 mAb，克隆 PMab-269（鼠 IgG，kappa）。PMab-269 通过流式细胞术和 Western blot 特异性检测过表达 seaPDPN 的中国仓鼠卵巢（CHO）-K1 细胞。此外，PMab-269 通过免疫组织化学清楚地区分了加利福尼亚海狮组织中的肺 I 型肺泡细胞、肾足细胞和结肠淋巴管内皮细胞。这些发现表明 PMab-269 可用于加利福尼亚海狮肺、肾和淋巴组织的病理生理学分析。
