Development of core-fucose-deficient humanized and chimeric anti-human podoplanin antibodies

Mika K. Kaneko, Tomokazu Ohishi, Takuro Nakamura, Hiroyuki Inoue, Junko Takei, Masato Sano, Teizo Asano, Yusuke Sayama, Hideki Hosono, Hiroyoshi Suzuki, Manabu Kawada, Yukinari Kato

Research output: Contribution to journalArticlepeer-review

Abstract

Podoplanin (PDPN), a 36-kDa type I transmembrane O-glycoprotein, is expressed in normal cells, including renal epithelial cells (podocytes), lymphatic endothelial cells, and pulmonary type I alveolar cells, and in cancer cells, including brain tumors and squamous cell lung carcinomas. PDPN activates platelet aggregation by binding to C-type lectin-like receptor-2 (CLEC-2) on platelets, and PDPN/CLEC-2 interaction facilitates blood/ lymphatic vessel separation. We previously produced an anti-human PDPN monoclonal antibody (mAb), clone NZ-1 (rat IgG2a, lambda) and its rat-human chimeric mAbs (NZ-8/NZ-12), which neutralize PDPN/CLEC-2 interactions and inhibit platelet aggregation and cancer metastasis. In this study, we first developed a humanized anti-human PDPN mAb, named as NZ-27. We further produced a core-fucose-deficient version of NZ-27, named as P1027 and a core-fucose-deficient version of NZ-12, named as NZ-12f. We investigated the binding affinity, antibody-dependent cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and antitumor activity of P1027 and NZ-12f. We demonstrated that the binding affinities of P1027 and NZ-12f against LN319 (a human glioblastoma cell line) are 1.1 · 10-8 and 3.9 · 10-9 M, respectively. ADCC reporter assays demonstrated that NZ-12f shows 1.5 times higher luminescence than P1027. Furthermore, NZ-12f showed 2.2 times higher ADCC than P1027, whereas both P1027 and NZ-12f showed high CDC activities against LN319 cells. Using LN319 xenograft models, P1027 and NZ-12f significantly reduced tumor development in an LN319 xenograft model compared with control human IgG. Treatment with P1027 and NZ-12f may be a useful therapy for patients with PDPN-expressing cancers.

Original languageEnglish
Pages (from-to)167-174
Number of pages8
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume39
Issue number5
DOIs
Publication statusPublished - 2020 Oct

Keywords

  • Humanized antibody
  • P1027
  • PDPN

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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