Yamamoto Haruto, Suzuki Hiroyuki, Tanaka Tomohiro, 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.
Biochem Biophys Rep. 2025 Jul 23;43:102170. doi: 10.1016/j.bbrep.2025.102170. eCollection 2025 Sep.
Podoplanin (PDPN) is a highly glycosylated type I transmembrane protein. PDPN expression is observed in various normal tissues, including lymphatic endothelial cells, kidney podocytes, and type I alveolar epithelial cells in the lungs. Monoclonal antibodies (mAbs) targeting PDPN across different animal species have facilitated the identification of PDPN-positive cells. To date, we have developed anti-PDPN mAbs for over 20 species. These antibodies suit various applications, including flow cytometry, immunoblotting, and immunohistochemistry. In this study, we generated an anti-hippopotamus PDPN (hipPDPN) mAb, PMab-322 (mouse IgG, kappa), using the Cell-Based Immunization and Screening (CBIS) method. PMab-322 exhibited strong reactivity to hipPDPN-overexpressed Chinese hamster ovary-K1 and demonstrated moderate affinity ( : 4.4 × 10 M) in a flow cytometry-based measurement. PMab-322 specifically recognizes hipPDPN but does not cross-react with PDPN from 23 other species. Furthermore, PMab-322 successfully detected hipPDPN in both immunoblotting and immunohistochemistry. These findings highlight the potential of PMab-322 for pathological analyses of hippopotamus-derived tissues.
血小板源性生长因子结合蛋白（PDPN）是一种高度糖基化的I型跨膜蛋白。在包括淋巴管内皮细胞、肾足细胞和肺I型肺泡上皮细胞在内的各种正常组织中都观察到PDPN的表达。针对不同动物物种的靶向PDPN的单克隆抗体（mAb）有助于识别PDPN阳性细胞。迄今为止，我们已经为20多种物种开发了抗PDPN单克隆抗体。这些抗体适用于各种应用，包括流式细胞术、免疫印迹和免疫组织化学。在本研究中，我们使用基于细胞的免疫和筛选（CBIS）方法制备了一种抗河马PDPN（hipPDPN）单克隆抗体PMab-322（小鼠IgG，κ）。PMab-322对过表达hipPDPN的中国仓鼠卵巢-K1细胞表现出强烈反应性，并在基于流式细胞术的测量中显示出中等亲和力（解离常数：4.4×10⁻⁸M）。PMab-322特异性识别hipPDPN，但不与其他23种物种的PDPN发生交叉反应。此外，PMab-322在免疫印迹和免疫组织化学中均成功检测到hipPDPN。这些发现突出了PMab-322在河马来源组织病理分析中的潜力。
