Abstract:
The giant panda (Ailuropoda melanoleuca) is one of the important species in worldwide animal conservation. Because it is essential to understand the disease of giant panda for conservation, histopathological analyses of tissues are important to understand the pathogenesis. However, monoclonal antibodies (mAbs) against giant panda-derived proteins are limited. Podoplanin (PDPN) is an essential marker of lung type I alveolar epithelial cells, kidney podocytes, and lymphatic endothelial cells. PDPN is also overexpressed in various human tumors, which are associated with poor prognosis. Here, an anti-giant panda PDPN (gpPDPN) mAb, PMab-314 (mouse IgG1, kappa) was established using the Cell-Based Immunization and Screening method. PMab-314 recognized N-terminal PA16-tagged gpPDPN-overexpressed Chinese hamster ovary-K1 cells (CHO/PA16-gpPDPN) in flow cytometry. The KD value of PMab-314 for CHO/PA16-gpPDPN was determined as 1.3 × 10-8 M. Furthermore, PMab-314 is useful for detecting gpPDPN in western blot analysis. These findings indicate that PMab-314 is a useful tool for the analyses of gpPDPN-expressed cells.
摘要:
大熊猫(Ailuropodamelanoleuca)是世界动物保护的重要物种之一。因为了解大熊猫的疾病对于保护至关重要,组织病理学分析对于理解发病机制很重要。然而,针对大熊猫来源蛋白质的单克隆抗体(mAb)有限。Podoplanin(PDPN)是肺I型肺泡上皮细胞的重要标志物,肾足细胞,和淋巴内皮细胞.PDPN在各种人类肿瘤中也过表达,这与预后不良有关。这里,抗大熊猫PDPN(gpPDPN)单克隆抗体,使用基于细胞的免疫和筛选方法建立PMab-314(小鼠IgG1,κ)。PMab-314在流式细胞术中识别N末端PA16标记的gpPDPN过表达的中国仓鼠卵巢K1细胞(CHO/PA16-gpPDPN)。CHO/PA16-gpPDPN的PMab-314的KD值被确定为1.3×10-8M。PMab-314可用于在蛋白质印迹分析中检测gpPDPN。这些发现表明PMab-314是分析gpPDPN表达的细胞的有用工具。