Characterization of anti-podoplanin monoclonal antibodies: critical epitopes for neutralizing the interaction between podoplanin and CLEC-2.

Abstract

Podoplanin (Aggrus) is a mucin-type sialoglycoprotein that is known as a useful marker for lymphatic endothelium and tumor-initiating cells (TICs). Interaction between podoplanin and C-type lectin-like receptor-2 (CLEC-2) is reported to be critical for podoplanin-induced platelet aggregation and cancer metastasis. Recently, several anti-human podoplanin antibodies have been created; however, these anti-podoplanin antibodies have not been well characterized. Five anti-podoplanin antibodies (NZ-1, D2-40, AB3, 18H5, and a rabbit polyclonal antibody) were investigated using ELISA, Western blot, and flow cytometry with synthesized podoplanin peptides and deletion mutants of recombinant podoplanin. The epitope of NZ-1 is platelet aggregation-stimulating (PLAG) domain-2/3; the epitope of D2-40, AB3, and 18H5 is PLAG1/2. The epitopes of D2-40 and AB3 are quite similar, although 18H5 is different from D2-40 and AB3. Using flow cytometric analysis, NZ-1 partially inhibited the interaction between podoplanin and CLEC-2, although other antibodies did not. In conclusion, the two most frequently used anti-podoplanin antibodies, D2-40 and AB3, have similar properties, although several studies have reported differences. NZ-1 neutralizes the interaction between podoplanin and CLEC-2, which may lead to the development of therapeutic antibodies against podoplanin-dependent cancer metastasis.

• Authors: Satoshi Ogasawara, Mika Kato Kaneko, Janet E Price, Yukinari Kato
• Journal: Hybridoma (2005) (Hybridoma (Larchmt)) Vol. 27 Issue 4 Pg. 259-67 (Aug 2008) ISSN: 1557-8348 [Electronic] United States
• PMID: 18707544 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• CLEC2B protein, human
• Epitopes
• Lectins, C-Type
• Membrane Glycoproteins
• PDPN protein, human
• Peptide Fragments

Topics

• Amino Acid Sequence
• Animals
• Antibodies, Monoclonal (immunology, pharmacology)
• CHO Cells
• Cells, Cultured
• Cricetinae
• Cricetulus
• Epitope Mapping
• Epitopes (immunology, metabolism)
• Humans
• Lectins, C-Type (metabolism)
• Membrane Glycoproteins (antagonists & inhibitors, immunology, metabolism)
• Molecular Sequence Data
• Neoplasms (immunology, pathology, therapy)
• Peptide Fragments (immunology, pharmacology)
• Protein Binding (drug effects)