Development of an anti-bear podoplanin monoclonal antibody PMab-247 for immunohistochemical analysis

Yoshikazu Furusawa; Junko Takei; Yusuke Sayama; Shinji Yamada; Mika K Kaneko; Yukinari Kato

doi:10.1016/j.bbrep.2019.100644

Development of an anti-bear podoplanin monoclonal antibody PMab-247 for immunohistochemical analysis

Biochem Biophys Rep. 2019 Apr 28:18:100644. doi: 10.1016/j.bbrep.2019.100644. eCollection 2019 Jul.

Authors

Yoshikazu Furusawa^{1

2}, Junko Takei¹, Yusuke Sayama¹, Shinji Yamada¹, Mika K Kaneko¹, Yukinari Kato^{1

2}

Affiliations

¹ Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, Japan.
² New Industry Creation Hatchery Center, Tohoku University, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, Japan.

Abstract

Sensitive and specific monoclonal antibodies (mAbs) targeting podoplanin (PDPN) are needed for immunohistochemical analyses using formalin-fixed paraffin-embedded tissues because PDPN is known as a lymphatic endothelial cell maker in pathology. Recently, we established anti-PDPN mAbs against many species, such as human, mouse, rat, rabbit, dog, cat, bovine, pig, horse, goat, tiger, alpaca, and Tasmanian devil. However, anti-bear PDPN (bPDPN) has not been established yet. In this study, we immunized mice with bPDPN-overexpressing Chinese hamster ovary (CHO)-K1 (CHO/bPDPN) cells, and screened mAbs against bPDPN using flow cytometry. One of the mAbs, PMab-247 (IgG₁, kappa), specifically detected CHO/bPDPN cells by flow cytometry and immunohistochemistry. Our findings suggest the potential usefulness of PMab-247 for the functional analyses of bPDPN.

Keywords: BAP, bovine aggrus/podoplanin; Bear podoplanin; DAB, 3,3′-diaminobenzidine tetrahydrochloride; ELISA, enzyme-linked immunosorbent assay; Immunohistochemistry; PBS, phosphate-buffered saline; PDPN; PDPN, podoplanin; PMab-247; mAb, monoclonal antibody.