Furusawa Yoshikazu, Yamada Shinji, Itai Shunsuke, Nakamura Takuro, Takei Junko, Sano Masato, Harada Hiroyuki, Fukui Masato, Kaneko Mika K, Kato Yukinari
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, Japan.
New Industry Creation Hatchery Center, Tohoku University, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan.
Biochem Biophys Rep. 2019 Apr 1;18:100631. doi: 10.1016/j.bbrep.2019.100631. eCollection 2019 Jul.
Monoclonal antibodies (mAbs) against not only human, mouse, and rat but also rabbit, dog, cat, bovine, pig, and horse podoplanins (PDPNs) have been established in our previous studies. PDPN is used as a lymphatic endothelial cell marker in pathological diagnoses. However, mAbs against Tasmanian devil PDPN (tasPDPN), which are useful for immunohistochemical analysis, remain to be developed. Herein, mice were immunized with tasPDPN-overexpressing Chinese hamster ovary (CHO)-K1 (CHO/tasPDPN) cells, and hybridomas producing mAbs against tasPDPN were screened using flow cytometry. One of the mAbs, PMab-233 (IgG, kappa), specifically detected CHO/tasPDPN cells by flow cytometry and recognized tasPDPN protein by western blotting. Furthermore, PMab-233 strongly detected CHO/tasPDPN cells by immunohistochemistry. These findings suggest that PMab-233 may be useful as a lymphatic endothelial cell marker of the Tasmanian devil.
在我们之前的研究中，已经制备出了不仅针对人、小鼠和大鼠，还针对兔、犬、猫、牛、猪和马的血小板内皮细胞黏附分子（PDPN）的单克隆抗体（mAb）。PDPN在病理诊断中用作淋巴管内皮细胞标记物。然而，针对袋獾PDPN（tasPDPN）的、可用于免疫组织化学分析的单克隆抗体仍有待开发。在此，用过量表达tasPDPN的中国仓鼠卵巢（CHO）-K1（CHO/tasPDPN）细胞免疫小鼠，并使用流式细胞术筛选产生抗tasPDPN单克隆抗体的杂交瘤。其中一种单克隆抗体PMab-233（IgG，κ）通过流式细胞术特异性检测CHO/tasPDPN细胞，并通过蛋白质印迹法识别tasPDPN蛋白。此外，PMab-233通过免疫组织化学强烈检测到CHO/tasPDPN细胞。这些发现表明，PMab-233可能作为袋獾淋巴管内皮细胞标记物有用。
