title: Affinity Maturation of Monoclonal Antibody 1E11 by Targeted Randomization in CDR3 Regions Optimizes Therapeutic Antibody Targeting of HER2-Positive Gastric Cancer.
abstract: Anti-HER2 murine monoclonal antibody 1E11 has strong and synergistic anti-tumor activity in HER2-overexpressing gastric cancer cells when used in combination with trastuzumab. We presently optimized this antibody for human therapeutics. First, the complementarity determining regions (CDRs) of the murine antibody were grafted onto human germline immunoglobulin variable genes. No difference in affinity and biological activity was observed between chimeric 1E11 (ch1E11) and humanized 1E11 (hz1E11). Next, affinity maturation of hz1E11 was performed by the randomization of CDR-L3 and H3 residues followed by stringent biopanning selection. Milder selection pressure favored the selection of more diverse clones, whereas higher selection stringency resulted in the convergence of the panning output to a smaller number of clones with improved affinity. Clone 1A12 had four amino acid substitutions in CDR-L3, and showed a 10-fold increase in affinity compared to the parental clone and increased potency in an in vitro anti-proliferative activity assay with HER2-overepxressing gastric cancer cells. Clone 1A12 inhibited tumor growth of NCI-N87 xenograft model with similar efficacy to trastuzumab alone, and the combination treatment of 1A12 and trastuzumab completely removed the established tumors. These results suggest that humanized and affinity matured monoclonal antibody 1A12 is a highly optimized molecule for future therapeutic development against HER2-positive tumors.
authors.affiliation: Therapeutic antibody research center, AbClon Inc., Seoul, Korea.
authors.forename: Bong-Kook
authors.initials: BK
authors.lastname: Ko
authors.affiliation: Therapeutic antibody research center, AbClon Inc., Seoul, Korea.
authors.forename: Soyoung
authors.initials: S
authors.lastname: Choi
authors.affiliation: Therapeutic antibody research center, AbClon Inc., Seoul, Korea.
authors.forename: Lei Guang
authors.initials: LG
authors.lastname: Cui
authors.affiliation: Therapeutic antibody research center, AbClon Inc., Seoul, Korea.
authors.forename: Young-Ha
authors.initials: YH
authors.lastname: Lee
authors.affiliation: Therapeutic antibody research center, AbClon Inc., Seoul, Korea.
authors.forename: In-Sik
authors.initials: IS
authors.lastname: Hwang
authors.affiliation: Therapeutic antibody research center, AbClon Inc., Seoul, Korea.
authors.forename: Kyu-Tae
authors.initials: KT
authors.lastname: Kim
authors.affiliation: Departments of Bioinspired Science and Life Science, Ewha Womans University, Seoul, Korea.
authors.forename: Hyunbo
authors.initials: H
authors.lastname: Shim
authors.affiliation: Therapeutic antibody research center, AbClon Inc., Seoul, Korea.
authors.forename: Jong-Seo
authors.initials: JS
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry / Q000276:immunology* / Q000627:therapeutic use*; D000915:Antibody Affinity; D045744:Cell Line, Tumor; D022801:Complementarity Determining Regions / Q000276:immunology*; D005260:Female; D018734:Genes, erbB-2; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013274:Stomach Neoplasms / Q000235:genetics / Q000628:therapy*
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D022801:Complementarity Determining Regions