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

IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry and Biophysics Reports Pub Date : 2025-07-23 DOI:10.1016/j.bbrep.2025.102170

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

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引用次数: 0

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_2a, 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 (K_D: 4.4 × 10⁻⁸ 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.

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PMab-322：一种新型多用途抗河马足planin单克隆抗体

Podoplanin （PDPN）是一种高度糖基化的I型跨膜蛋白。PDPN在各种正常组织中均有表达，包括淋巴内皮细胞、肾足细胞和肺I型肺泡上皮细胞。针对不同动物物种的PDPN的单克隆抗体（mab）促进了PDPN阳性细胞的鉴定。迄今为止，我们已经针对20多个物种开发了抗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）方法生成了抗河马PDPN （hipPDPN）单抗，PMab-322（小鼠IgG2a， kappa）。在基于流式细胞术的测量中，PMab-322对过表达的中国仓鼠卵巢k1表现出很强的反应性，并表现出中等亲和力（KD: 4.4 × 10−8 M）。PMab-322特异性识别hipdpn，但不与其他23种PDPN交叉反应。此外，PMab-322在免疫印迹和免疫组织化学中成功检测到hipdpn。这些发现突出了PMab-322用于河马源性组织病理分析的潜力。

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来源期刊

Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

191

审稿时长

59 days

期刊介绍： Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.