Development of an Anti-Sheep Podoplanin Monoclonal Antibody PMab-256 for Immunohistochemical Analysis of Lymphatic Endothelial Cells

Yukinari Kato, Yoshikazu Furusawa, Masato Sano, Junko Takei, Takuro Nakamura, Miyuki Yanaka, Saki Okamoto, Saori Handa, Yu Komatsu, Teizo Asano, Yusuke Sayama, Mika K. Kaneko

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Sensitive and specific monoclonal antibodies (mAbs) targeting podoplanin (PDPN) are needed for immunohistochemical analyses as a marker for lymphatic endothelial cells. We recently have developed anti-PDPN mAbs against many species, including human, mouse, rat, rabbit, dog, cat, bovine, pig, Tasmanian devil, alpaca, tiger, whale, goat, horse, and bear. However, anti-sheep PDPN (sPDPN) has not yet been established. In this study, we used the Cell-Based Immunization and Screening method for the development of anti-sPDPN mAbs. RAP14 tag was added to N-terminus of sPDPN, and anti-RAP14 tag mAb (PMab-2) was used to detect the expression level of sPDPN in flow cytometry and western blot. We immunized mice with sPDPN-overexpressing Chinese hamster ovary (CHO)-K1 (CHO/sPDPN) cells and screened mAbs against sPDPN using flow cytometry. One of the mAbs, PMab-256 (IgG1, kappa), specifically detected CHO/sPDPN cells by flow cytometry and western blot. Furthermore, PMab-256 stained type I alveolar cells of lung, renal glomerulus and Bowman's capsule, and lymphatic endothelial cells of lung and colon. Our findings suggest the potential usefulness of PMab-256 for the functional analyses of sPDPN.

Original languageEnglish
Pages (from-to)82-90
Number of pages9
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume39
Issue number3
DOIs
Publication statusPublished - 2020 Jun

Keywords

  • monoclonal antibody
  • podoplanin
  • sPDPN
  • sheep

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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