title: Effective therapeutic approach for head and neck cancer by an engineered minibody targeting the EGFR receptor.
abstract: Cetuximab, a chimeric monoclonal antibody developed for targeting the Epidermal Growth Factor Receptor (EGFR), has been intensively used to treat cancer patients with metastatic colorectal cancer and head and neck cancer. Intact immunoglobulin G (IgG) antibody like cetuximab, however, has some limitations such as high production cost and low penetration rate from vasculature into solid tumor mass due to its large size. In attempt to overcome these limitations, we engineered cetuximab to create single chain variable fragments (scFv-CH3; Minibody) that were expressed in bacterial system. Among three engineered minibodies, we found that MI061 minibody, which is composed of the variable heavy (VH) and light (VL) region joined by an 18-residue peptide linker, displays higher solubility and better extraction properties from bacterial lysate. In addition, we validated that purified MI061 significantly interferes ligand binding to EGFR and blocks EGFR's phosphorylation. By using a protein microarray composed of 16,368 unique human proteins covering around 2,400 plasma membrane associated proteins such as receptors and channels, we also demonstrated that MI061 only recognizes the EGFR but not other proteins as compared with cetuximab. These results indicated that engineered MI061 retains both binding specificity and affinity of cetuximab for EGFR. Although it had relatively short half-life in serum, it was shown to be highly significant anti-tumor effect by inhibiting ERK pathway in A431 xenograft model. Taken together, our present study provides compelling evidence that engineered minibody is more effective and promising agent for in vivo targeting of solid tumors.
authors.affiliation: Department of Bio-Engineering, Life Science RD Center, Sinil Pharmaceutical Co., Seongnam, South Korea.
authors.forename: Young Pil
authors.initials: YP
authors.lastname: Kim
authors.affiliation: Department of Physiology, Ajou University School of Medicine, Suwon, South Korea.
authors.forename: Dongsun
authors.initials: D
authors.lastname: Park
authors.affiliation: Department of Physiology, Ajou University School of Medicine, Suwon, South Korea; Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea.
authors.forename: Jae Jin
authors.initials: JJ
authors.lastname: Kim
authors.forename: Won-Jae
authors.initials: WJ
authors.lastname: Chi
authors.affiliation: Department of Bio-Engineering, Life Science RD Center, Sinil Pharmaceutical Co., Seongnam, South Korea.
authors.forename: Won-Jae
authors.initials: WJ
authors.lastname: Choi
authors.affiliation: Department of Bio-Engineering, Life Science RD Center, Sinil Pharmaceutical Co., Seongnam, South Korea.
authors.forename: Sun Hee
authors.initials: SH
authors.lastname: Lee
authors.affiliation: Department of Physiology, Ajou University School of Medicine, Suwon, South Korea; Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea.
authors.forename: Seo Yun
authors.initials: SY
authors.lastname: Lee
authors.affiliation: Department of Physiology, Ajou University School of Medicine, Suwon, South Korea; Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea.
authors.forename: Soyeon
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Physiology, Ajou University School of Medicine, Suwon, South Korea; Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea.
authors.forename: Jee Min
authors.initials: JM
authors.lastname: Chung
authors.affiliation: Department of Physiology, Ajou University School of Medicine, Suwon, South Korea.
authors.forename: Jinseon
authors.initials: J
authors.lastname: Jeon
authors.affiliation: Department of Neuroscience, Kyung Hee University School of Medicine, Seoul, South Korea.
authors.forename: Byoung Dae
authors.initials: BD
authors.lastname: Lee
authors.affiliation: Division of Pharmacology, Department of Molecular Cell Biology, Samsung Biomedical Research Institute, SungKyunKwan University School of Medicine, Suwon, South Korea.
authors.forename: Joo-Ho
authors.initials: JH
authors.lastname: Shin
authors.affiliation: Well Aging Research Center, Samsung Advanced Institute of Technology (SAIT), Suwon, South Korea.
authors.forename: Yun-Il
authors.initials: YI
authors.lastname: Lee
authors.affiliation: Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea; Department of Biochemistry, Ajou University School of Medicine, Suwon, South Korea.
authors.forename: Hyeseong
authors.initials: H
authors.lastname: Cho
authors.affiliation: Department of Bio-Engineering, Life Science RD Center, Sinil Pharmaceutical Co., Seongnam, South Korea.
authors.forename: Jeong-Min
authors.initials: JM
authors.lastname: Lee
authors.affiliation: Department of Physiology, Ajou University School of Medicine, Suwon, South Korea; Department of Biomedical Sciences, The Graduate School, Ajou University, Suwon, South Korea.
authors.forename: Ho Chul
authors.initials: HC
authors.lastname: Kang
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000235:genetics*; D000918:Antibody Specificity; D001483:Base Sequence; D045744:Cell Line, Tumor; D000068818:Cetuximab; D066246:ErbB Receptors / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D006258:Head and Neck Neoplasms / Q000188:drug therapy* / Q000276:immunology / Q000473:pathology; D006801:Humans; D008297:Male; D051379:Mice; D058990:Molecular Targeted Therapy; D015202:Protein Engineering; D015398:Signal Transduction / Q000276:immunology; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology / Q000627:therapeutic use*; D023041:Xenograft Model Antitumor Assays
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D057127:Single-Chain Antibodies; D066246:ErbB Receptors; D000068818:Cetuximab