Robo4 is an effective tumor endothelial marker for antibody-drug conjugates based on the rapid isolation of the anti-Robo4 cell-internalizing antibody.
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.
Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan.
Mai
M
Yoshikawa
Yohei
Y
Mukai
Yoshiaki
Y
Okada
Yuki
Y
Tsumori
Shin-ichi
S
Tsunoda
Yasuo
Y
Tsutsumi
William C
WC
Aird
Yasuo
Y
Yoshioka
Naoki
N
Okada
Takefumi
T
Doi
Shinsaku
S
Nakagawa
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000493:pharmacokinetics*; D000918:Antibody Specificity; D001692:Biological Transport / Q000276:immunology; D014408:Biomarkers, Tumor / Q000276:immunology / Q000378:metabolism; D002461:Cell Line, Transformed; D015195:Drug Design; D005260:Female; D057166:High-Throughput Screening Assays; D006801:Humans; D018796:Immunoconjugates / Q000276:immunology / Q000493:pharmacokinetics*; D008545:Melanoma / Q000098:blood supply / Q000276:immunology / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D009389:Neovascularization, Pathologic / Q000276:immunology / Q000628:therapy; D009419:Nerve Tissue Proteins / Q000276:immunology* / Q000378:metabolism*; D019151:Peptide Library; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic / Q000276:immunology* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism; D012878:Skin Neoplasms / Q000098:blood supply / Q000276:immunology / Q000628:therapy*; D014018:Tissue Distribution; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D018796:Immunoconjugates; D009419:Nerve Tissue Proteins; D019151:Peptide Library; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D011994:Recombinant Proteins; C540792:Robo4 protein, mouse; D040301:Vascular Endothelial Growth Factor Receptor-2