Development of sensitive monoclonal antibody PMab-2 against rat podoplanin

Hiroharu Oki, Ryusuke Honma, Satoshi Ogasawara, Yuki Fujii, Xing Liu, Michiaki Takagi, Mika K. Kaneko, Yukinari Kato

Research output: Contribution to journalArticlepeer-review

46 Citations (Scopus)

Abstract

Podoplanin (PDPN) is a platelet aggregation-inducing factor, which is known as an endogenous ligand of C-type lectin-like receptor-2 (CLEC-2). PDPN is also expressed in several normal tissues, such as lung type I alveolar cells and kidney podocytes. Although many monoclonal antibodies (MAbs) against human PDPN (hPDPN) or mouse PDPN (mPDPN) have been established, anti-rat PDPN (rPDPN) MAbs, especially against platelet aggregation-stimulating (PLAG) domain (29-54 amino acids) of rPDPN, have not been developed. Therefore, functional analysis of rPDPN in normal tissues has been limited. Here, we immunized mice with rPDPN peptides (38-51 amino acids) and developed a novel mouse anti-rPDPN MAb, PMab-2 (IgG1, kappa), which possesses high affinity compared with anti-hPDPN or mPDPN MAbs. The KD of PMab-2 was determined to be 5.9 × 10-10 M. PMab-2 is useful, not only in flow cytometry and Western blot analysis against endogenous rPDPN, which is expressed in rat dermal fibroblast, but also in immunohistochemistry against normal tissues. PMab-2 showed extraordinarily high sensitivity in immunohistochemistry, indicating that PMab-2 is very advantageous for functional analysis of rPDPN.

Original languageEnglish
Pages (from-to)396-403
Number of pages8
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume34
Issue number6
DOIs
Publication statusPublished - 2015 Dec 1

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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