使用抗山羊组织的免疫组织化学方法对抗山羊血小板反应蛋白-1结构域7单克隆抗体PMab-235的特性进行鉴定。
Characterization of Anti-Goat Podoplanin Monoclonal Antibody PMab-235 Using Immunohistochemistry Against Goat Tissues.
作者信息
Takei Junko, Itai Shunsuke, Harada Hiroyuki, Furusawa Yoshikazu, Miwa Takashi, Fukui Masato, Nakamura Takuro, Sano Masato, Sayama Yusuke, Yanaka Miyuki, Handa Saori, Hisamatsu Kayo, Nakamura Yoshimi, Yamada Shinji, Kaneko Mika K, Kato Yukinari
机构信息
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Department of Oral and Maxillofacial Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan.
出版信息
Monoclon Antib Immunodiagn Immunother. 2019 Oct;38(5):213-219. doi: 10.1089/mab.2019.0022. Epub 2019 Aug 12.
Podoplanin (PDPN)/T1alpha is expressed on lymphatic endothelial cells, type I alveolar cells of the lungs, and podocytes of the kidney. PDPN possesses three platelet aggregation-stimulating (PLAG) domains (PLAG1, PLAG2, and PLAG3) of the N-terminus and the PLAG-like domains (PLDs). We previously reported an anti-goat PDPN (gPDPN) monoclonal antibody (mAb), PMab-235, which was developed using the Cell-Based Immunization and Screening (CBIS) method. PMab-235 is very useful in flow cytometry, Western blotting, and immunohistochemical analyses; however, the binding epitope of PMab-235 remains to be elucidated. In this study, we investigated the epitopes of PMab-235 using enzyme-linked immunosorbent assay and immunohistochemistry. The results revealed that the critical epitope of PMab-235 produced by CBIS method is Arg75, Leu78, and Pro79 of gPDPN, which is included in PLD. The findings of our study can be applied to the production of more functional anti-gPDPN mAbs.
血小板源性生长因子（PDPN）/T1α在淋巴管内皮细胞、肺的I型肺泡细胞和肾足细胞中表达。PDPN在N端具有三个血小板聚集刺激（PLAG）结构域（PLAG1、PLAG2和PLAG3）以及类PLAG结构域（PLD）。我们之前报道了一种抗山羊PDPN（gPDPN）单克隆抗体（mAb）PMab - 235，它是采用基于细胞的免疫和筛选（CBIS）方法研制的。PMab - 235在流式细胞术、蛋白质印迹和免疫组织化学分析中非常有用；然而，PMab - 235的结合表位仍有待阐明。在本研究中，我们使用酶联免疫吸附测定和免疫组织化学研究了PMab - 235的表位。结果显示，通过CBIS方法产生的PMab - 235的关键表位是gPDPN的第75位精氨酸、第78位亮氨酸和第79位脯氨酸，它们包含在PLD中。我们的研究结果可应用于生产更多功能性抗gPDPN单克隆抗体。
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