AccScience Publishing / MI / Online First / DOI: 10.36922/MI025160033
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Development of novel anti-rhinoceros podoplanin monoclonal antibodies for flow cytometry, western blotting, and immunohistochemistry

Shiori Fujisawa1 ,  Rena Ubukata1 ,  Hiroyuki Suzuki1* ,  Tomohiro Tanaka1 ,  Airi Nomura1 ,  Keisuke Shinoda1 ,  Takuya Nakamura1 ,  Hiroyuki Satofuka1 ,  Guanjie Li1 ,  Mika K. Kaneko1 ,  Yukinari Kato1*
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1 Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan
MI, 025160033 https://doi.org/10.36922/MI025160033
Received: 14 April 2025 | Revised: 16 July 2025 | Accepted: 1 September 2025 | Published online: 19 September 2025
© 2025 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Podoplanin (PDPN) is a type I transmembrane glycoprotein characterized by a highly glycosylated extracellular domain, a single transmembrane domain, and a short intracellular tail. PDPN is physiologically expressed in various normal tissues and cell types, including pulmonary type I alveolar epithelial cells, renal podocytes, and lymphatic endothelial cells. In addition to its physiological roles, PDPN has emerged as a valuable biomarker in cancer research. Consequently, the development of anti-PDPN monoclonal antibodies (mAbs) across a range of animal species has facilitated the identification and characterization of PDPN-expressing cells. To date, we have generated anti-PDPN mAbs for over 20 species, suitable for applications, such as flow cytometry, Western blotting, and immunohistochemistry. In the present study, we established two anti-rhinoceros PDPN (rhiPDPN) mAbs—PMab-315 (mouse IgG2a, kappa) and PMab-324 (mouse IgG2b, kappa)—using the Cell-Based Immunization and Screening method. Both mAbs bound to rhiPDPN-overexpressed Chinese hamster ovary K1 (CHO/rhiPDPN) and exhibited an identical KD value (4.5 × 10⁻⁸ M) in a flow cytometry-based assay. Furthermore, PMab-315 and PMab-324 effectively detected rhiPDPN in both Western blot and immunohistochemical analyses. These findings highlight the potential utility of PMab-315 and PMab-324 as robust tools for the pathological investigation of rhinoceros-derived tissues and cells.

Keywords
Rhinoceros podoplanin
Monoclonal antibody
CBIS method
Flow cytometry
Funding
This research was supported in part by the Japan Agency for Medical Research and Development (AMED) under grant numbers: JP25am0521010 (to Y.K.), JP25am0521010 (to Y.K.), JP24ama221339 (to Y.K.), JP24bm1123027 (to Y.K.), and JP24ck0106730 (to Y.K.), and by the Japan Society for the Promotion of Science (JSPS) Grants-in-Aid for Scientific Research (KAKENHI) under grant numbers: 22K06995 (to H.Suzuki), 24K18268 (to T.T.), 24K11652 (to H.Satofuka), and 22K07224 (to Y.K.).
Conflict of interest
The authors declare that they have no competing interests.
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Microbes & Immunity, Electronic ISSN: 3029-2883 Print ISSN: 3041-0886, Published by AccScience Publishing
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