抗猪血小板内皮细胞黏附分子单克隆抗体PMab-213的表位图谱分析
Epitope Mapping of Antipig Podoplanin Monoclonal Antibody PMab-213.
作者信息
Yamada Shinji, Itai Shunsuke, Furusawa Yoshikazu, Kaneko Mika K, Kato Yukinari
机构信息
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
New Industry Creation Hatchery Center, Tohoku University, Sendai, Japan.
出版信息
Monoclon Antib Immunodiagn Immunother. 2019 Oct;38(5):224-229. doi: 10.1089/mab.2019.0023. Epub 2019 Aug 29.
Podoplanin (PDPN), a type I transmembrane sialoglycoprotein, is expressed on lymphatic endothelial cells, podocytes of the kidneys, and type I alveolar cells of the lungs. PDPN, a platelet aggregation-inducing factor, comprises three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) in the N-terminus and PLAG-like domains in the middle of the PDPN protein. We have previously reported a mouse antipig PDPN (pPDPN) monoclonal antibody (mAb) clone, PMab-213, which was developed using the Cell-Based Immunization and Screening (CBIS) method. PMab-213 is very useful in flow cytometry, Western blotting, and immunohistochemical analyses; however, the binding epitope of PMab-213, which was developed by CBIS method, remains to be elucidated. Therefore, this study aimed to investigate the epitope of PMab-213 using enzyme-linked immunosorbent assay, flow cytometry, and immunohistochemical analyses. The results revealed that the critical epitopes of PMab-213 are Pro53, Arg54, Arg56, and Tyr60 of pPDPN.
血小板内皮细胞黏附分子（Podoplanin，PDPN）是一种I型跨膜唾液酸糖蛋白，在淋巴管内皮细胞、肾足细胞和肺I型肺泡细胞中表达。PDPN作为一种血小板聚集诱导因子，在其N端包含三个血小板聚集刺激（PLAG）结构域（PLAG1、PLAG2和PLAG3），在PDPN蛋白中部包含类PLAG结构域。我们之前报道过一种小鼠抗猪PDPN（pPDPN）单克隆抗体（mAb）克隆体PMab - 213，它是采用基于细胞的免疫和筛选（CBIS）方法研制的。PMab - 213在流式细胞术、蛋白质免疫印迹和免疫组织化学分析中非常有用；然而，通过CBIS方法研制的PMab - 213的结合表位仍有待阐明。因此，本研究旨在利用酶联免疫吸附测定、流式细胞术和免疫组织化学分析来研究PMab - 213的表位。结果显示，PMab - 213的关键表位是pPDPN的Pro53、Arg54、Arg56和Tyr60。
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