Phage-derived fully human monoclonal antibody fragments to human vascular endothelial growth factor-C block its interaction with VEGF receptor-2 and 3.

Abstract

Vascular endothelial growth factor C (VEGF-C) is a key mediator of lymphangiogenesis, acting via its receptors VEGF-R2 and VEGF-R3. High expression of VEGF-C in tumors correlates with increased lymphatic vessel density, lymphatic vessel invasion, sentinel lymph node metastasis and poor prognosis. Recently, we found that in a chemically induced skin carcinoma model, increased VEGF-C drainage from the tumor enhanced lymphangiogenesis in the sentinel lymph node and facilitated metastatic spread of cancer cells via the lymphatics. Hence, interference with the VEGF-C/VEGF-R3 axis holds promise to block metastatic spread, as recently shown by use of a neutralizing anti-VEGF-R3 antibody and a soluble VEGF-R3 (VEGF-C/D trap). By antibody phage-display, we have developed a human monoclonal antibody fragment (single-chain Fragment variable, scFv) that binds with high specificity and affinity to the fully processed mature form of human VEGF-C. The scFv binds to an epitope on VEGF-C that is important for receptor binding, since binding of the scFv to VEGF-C dose-dependently inhibits the binding of VEGF-C to VEGF-R2 and VEGF-R3 as shown by BIAcore and ELISA analyses. Interestingly, the variable heavy domain (V(H)) of the anti-VEGF-C scFv, which contains a mutation typical for camelid heavy chain-only antibodies, is sufficient for binding VEGF-C. This reduced the size of the potentially VEGF-C-blocking antibody fragment to only 14.6 kDa. Anti-VEGF-C V(H)-based immunoproteins hold promise to block the lymphangiogenic activity of VEGF-C, which would present a significant advance in inhibiting lymphatic-based metastatic spread of certain cancer types.

• Authors: Matthias Rinderknecht, Alessandra Villa, Kurt Ballmer-Hofer, Dario Neri, Michael Detmar
• Journal: PloS one (PLoS One) Vol. 5 Issue 8 Pg. e11941 (Aug 02 2010) ISSN: 1932-6203 [Electronic] United States
• PMID: 20689828 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• Immunoglobulin Heavy Chains
• Peptide Library
• Single-Chain Antibodies
• Vascular Endothelial Growth Factor C
• Vascular Endothelial Growth Factor Receptor-2
• Vascular Endothelial Growth Factor Receptor-3

Topics

• Amino Acid Sequence
• Antibodies, Monoclonal (chemistry, genetics, immunology, metabolism)
• Antibody Affinity
• Antibody Specificity
• Enzyme-Linked Immunosorbent Assay
• Humans
• Hydrophobic and Hydrophilic Interactions
• Immunoglobulin Heavy Chains (immunology)
• Models, Molecular
• Molecular Sequence Data
• Mutation
• Peptide Library
• Protein Binding (immunology)
• Protein Multimerization
• Protein Stability
• Protein Structure, Quaternary
• Single-Chain Antibodies (chemistry, genetics, immunology, metabolism)
• Vascular Endothelial Growth Factor C (chemistry, immunology, metabolism)
• Vascular Endothelial Growth Factor Receptor-2 (metabolism)
• Vascular Endothelial Growth Factor Receptor-3 (metabolism)