Abstract
Monoclonal antibodies (mAbs) that are internalized into cells are a current focus in the development of antibody-drug conjugates (ADCs). We describe a phage display-based high-throughput screening system to rapidly isolate cell-internalizing mAbs. We simultaneously examined the cell-internalizing activities of several hundred independent mAbs and successfully isolated cell-internalizing mAbs against the tumor endothelial markers Roundabout homolog 4 (Robo4) and vascular endothelial growth factor receptor 2 (VEGFR2). Tumor accumulation of mAbs with high cell-internalizing activity was significantly higher than that of mAbs with low cell-internalizing activity. Furthermore, the antitumor effects of ADCs of mAbs with high cell-internalizing activity were significantly stronger than those of mAbs with low cell-internalizing activity. Although anti-VEGFR2 therapy caused a significant loss of body weight, anti-Robo4 therapy did not. These findings indicate that cell-internalizing activity plays an important role in the biodistribution and therapeutic effects of ADCs. Further, Robo4 can be an effective marker for tumor vascular targeting.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal / immunology
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Antibodies, Monoclonal / pharmacokinetics*
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Antibody Specificity
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Biological Transport / immunology
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Biomarkers, Tumor / immunology
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Biomarkers, Tumor / metabolism
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Cell Line, Transformed
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Drug Design
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Female
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High-Throughput Screening Assays
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Humans
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Immunoconjugates / immunology
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Immunoconjugates / pharmacokinetics*
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Melanoma / blood supply
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Melanoma / immunology
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Melanoma / therapy*
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Mice
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Mice, Inbred C57BL
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Neovascularization, Pathologic / immunology
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Neovascularization, Pathologic / therapy
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Nerve Tissue Proteins / immunology*
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Nerve Tissue Proteins / metabolism*
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Peptide Library
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Receptors, Cell Surface
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Receptors, Immunologic / immunology*
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Receptors, Immunologic / metabolism*
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Recombinant Proteins / immunology
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Recombinant Proteins / metabolism
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Skin Neoplasms / blood supply
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Skin Neoplasms / immunology
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Skin Neoplasms / therapy*
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Tissue Distribution
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Vascular Endothelial Growth Factor Receptor-2 / immunology
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Vascular Endothelial Growth Factor Receptor-2 / metabolism
Substances
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Antibodies, Monoclonal
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Biomarkers, Tumor
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Immunoconjugates
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Nerve Tissue Proteins
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Peptide Library
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Receptors, Cell Surface
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Receptors, Immunologic
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Recombinant Proteins
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Robo4 protein, mouse
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Vascular Endothelial Growth Factor Receptor-2