title: Generation of Potent Anti-Vascular Endothelial Growth Factor Neutralizing Antibodies from Mouse Phage Display Library for Cancer Therapy.
abstract: Vascular endothelial growth factor (VEGF) is an important stimulator for angiogenesis in solid tumors. Blocking VEGF activity is an effective therapeutic strategy to inhibit tumor growth and metastasis. Avastin, a humanized monoclonal antibody recognizes VEGF, has been approved by the US Food and Drug Administration. To generate potential VEGF-recognizing antibodies with better tumor regression ability than that of Avastin, we have designed a systematic antibody selection plan. From mice immunized with recombinant human VEGF, we generated three phage display libraries, scFv-M13KO7, Fab-M13KO7, and scFv-Hyperphage, in single-chain Fv (scFv) or Fab format, displayed using either M13KO7 helper phage or Hyperphage. Solid-phase and solution-phase selection strategies were then applied to each library, generating six panning combinations. A total of sixty-four antibodies recognizing VEGF were obtained. Based on the results of epitope mapping, binding affinity, and biological functions in tumor inhibition, eight antibodies were chosen to examine their abilities in tumor regression in a mouse xenograft model using human COLO 205 cancer cells. Three of them showed improvement in the inhibition of tumor growth (328%-347% tumor growth ratio (% of Day 0 tumor volume) on Day 21 vs. 435% with Avastin). This finding suggests a potential use of these three antibodies for VEGF-targeted therapy.
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. alienwhale@yahoo.com.tw.
authors: Yan-Da
authors: YD
authors: Lai
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. liannyuyu@dcb.org.tw.
authors: Yen-Yu
authors: YY
authors: Wu
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. yi_jiue@dcb.org.tw.
authors: Yi-Jiue
authors: YJ
authors: Tsai
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. yisantsai@dcb.org.tw.
authors: Yi-San
authors: YS
authors: Tsai
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. yuying1982@gmail.com.
authors: Yu-Ying
authors: YY
authors: Lin
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. lucanus@dcb.org.tw.
authors: Szu-Liang
authors: SL
authors: Lai
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. bchy@dcb.org.tw.
authors: Chao-Yang
authors: CY
authors: Huang
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. ziltoucan@dcb.org.tw.
authors: Ying-Yung
authors: YY
authors: Lok
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. cyhu@dcb.org.tw.
authors: Chih-Yung
authors: CY
authors: Hu
authors: Department of Protein Engineering, Development Center for Biotechnology, New Taipei City 22180, Taiwan. jslai@obipharma.com.
authors: Jiann-Shiun
authors: JS
authors: Lai
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D057134:Antibodies, Neutralizing / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D000915:Antibody Affinity / Q000276:immunology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D061505:Cell Surface Display Techniques; D004195:Disease Models, Animal; D018604:Epitope Mapping; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D019151:Peptide Library; D010766:Phosphorylation; D011485:Protein Binding; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000037:antagonists & inhibitors* / Q000276:immunology; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D023041:Xenograft Model Antitumor Assays
chemical_list: D020533:Angiogenesis Inhibitors; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D019151:Peptide Library; D057127:Single-Chain Antibodies; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Fab antibody fragment; angiogenesis; phage display; single chain Fv antibody fragment (scFv); vascular endothelial growth factor (VEGF)