Furusawa Yoshikazu, Yamada Shinji, Itai Shunsuke, Nakamura Takuro, Yanaka Miyuki, 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 Feb 2;18:100616. doi: 10.1016/j.bbrep.2019.01.009. eCollection 2019 Jul.
Monoclonal antibodies (mAbs) against human, mouse, rat, rabbit, dog, cat, and bovine podoplanin (PDPN), a lymphatic endothelial cell marker, have been established in our previous studies. However, mAbs against horse PDPN (horPDPN), which are useful for immunohistochemical analysis, remain to be developed. In the present study, mice were immunized with horPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/horPDPN), and hybridomas producing mAbs against horPDPN were screened using flow cytometry. One of the mAbs, PMab-219 (IgG, kappa), specifically detected CHO/horPDPN cells via flow cytometry and recognized horPDPN protein using Western blotting. Furthermore, PMab-219 strongly stained CHO/horPDPN via immunohistochemistry. These findings suggest that PMab-219 is useful for investigating the function of horPDPN.
在我们之前的研究中，已经制备了针对人、小鼠、大鼠、兔、狗、猫和牛的淋巴管内皮细胞标志物血小板内皮细胞黏附分子（PDPN）的单克隆抗体（mAb）。然而，对于免疫组织化学分析有用的抗马PDPN（horPDPN）单克隆抗体仍有待开发。在本研究中，用过量表达horPDPN的中国仓鼠卵巢（CHO）-K1细胞（CHO/horPDPN）免疫小鼠，并使用流式细胞术筛选产生抗horPDPN单克隆抗体的杂交瘤。其中一种单克隆抗体PMab-219（IgG，κ）通过流式细胞术特异性检测CHO/horPDPN细胞，并使用蛋白质印迹法识别horPDPN蛋白。此外，PMab-219通过免疫组织化学强烈染色CHO/horPDPN。这些发现表明PMab-219可用于研究horPDPN的功能。
