Department of Molecular Pharmacology and Tohoku University Graduate School of Medicine, Sendai, Japan.
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Monoclon Antib Immunodiagn Immunother. 2022 Aug;41(4):194-201. doi: 10.1089/mab.2022.0007. Epub 2022 Aug 2.
Podoplanin (PDPN) is an essential marker of lung type I alveolar cells, kidney podocytes, and lymphatic endothelial cells. Monoclonal antibodies (mAbs) that can specifically recognize PDPN in immunohistochemistry are important to analyze the development of tissues and the pathogenesis of diseases, including cancers. We have developed anti-PDPN mAbs against many animal species; however, mAbs that can recognize elephant-derived membrane proteins and distinguish the specific cell types in immunohistochemistry are limited. In this study, a novel anti-elephant PDPN (elePDPN) mAb, PMab-295 (IgG, kappa), was established using the peptide immunization method. PMab-295 recognized both elePDPN-overexpressed Chinese hamster ovary (CHO)-K1 cells and endogenous elePDPN-expressed LACF-NaNaI cells by flow cytometry and western blotting. Kinetic analyses using flow cytometry showed that the of PMab-295 for CHO/elePDPN was 1.5 × 10 M. Furthermore, PMab-295 detected elePDPN-expressing cells using immunohistochemistry. These results showed the usefulness of PMab-295 to investigate the molecular function of elePDPN and the pathogenesis of diseases.
波形蛋白（PDPN）是肺 I 型肺泡细胞、肾脏足细胞和淋巴管内皮细胞的重要标志物。能够在免疫组织化学中特异性识别 PDPN 的单克隆抗体（mAbs）对于分析组织发育和疾病发病机制（包括癌症）非常重要。我们已经针对许多动物物种开发了抗 PDPN 的 mAbs；然而，能够识别大象来源的膜蛋白并在免疫组织化学中区分特定细胞类型的 mAbs 是有限的。在这项研究中，我们使用肽免疫法建立了一种新型抗象 PDPN（elePDPN）mAb，即 PMab-295（IgG，kappa）。PMab-295 通过流式细胞术和 Western blot 识别过表达 elePDPN 的中国仓鼠卵巢（CHO）-K1 细胞和内源性表达 elePDPN 的 LACF-NaNaI 细胞。使用流式细胞术进行的动力学分析表明，PMab-295 与 CHO/elePDPN 的亲和力为 1.5×10-8M。此外，PMab-295 通过免疫组织化学检测 elePDPN 表达细胞。这些结果表明 PMab-295 可用于研究 elePDPN 的分子功能和疾病发病机制。
