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PMab-322:一种适用于多种用途的新型抗河马血小板反应蛋白单克隆抗体。

PMab-322: a novel anti-hippopotamus podoplanin monoclonal antibody for multiple applications.

作者信息

Yamamoto Haruto, Suzuki Hiroyuki, Tanaka Tomohiro, Kaneko Mika K, Kato Yukinari

机构信息

Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, Japan.

出版信息

Biochem Biophys Rep. 2025 Jul 23;43:102170. doi: 10.1016/j.bbrep.2025.102170. eCollection 2025 Sep.

Abstract

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.

摘要

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。

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