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

## TL;DR

A new monoclonal antibody, PMab-322, was developed to detect podoplanin in hippopotamus tissues, aiding in their pathological analysis.

## Contribution

Development of a novel monoclonal antibody specific to hippopotamus podoplanin with applications in multiple assays.

## Key findings

- PMab-322 specifically recognizes hippopotamus podoplanin without cross-reacting with 23 other species.
- PMab-322 is effective in flow cytometry, immunoblotting, and immunohistochemistry for detecting hippopotamus podoplanin.
- The antibody has moderate affinity (KD: 4.4 × 10−8 M) and is suitable for pathological analyses of hippopotamus-derived tissues.

## 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 IgG2a, 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 (KD: 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.

•A novel anti-hippopotamus podoplanin (hipPDPN) monoclonal antibody, PMab-322 was developed by the Cell-Based Immunization and Screening method.•PMab-322 detects hipPDPN by flow cytometry.•PMab-322 is also suitable for detecting hipPDPN in immunoblotting and immunohistochemistry.•PMab-322 could contribute to the pathological analysis of hippopotamus tissues.

A novel anti-hippopotamus podoplanin (hipPDPN) monoclonal antibody, PMab-322 was developed by the Cell-Based Immunization and Screening method.

PMab-322 detects hipPDPN by flow cytometry.

PMab-322 is also suitable for detecting hipPDPN in immunoblotting and immunohistochemistry.

PMab-322 could contribute to the pathological analysis of hippopotamus tissues.