Highly Effective Recombinant Format of a Humanized IgG-like Bispecific Antibody for Cancer Immunotherapy with Retargeting of Lymphocytes to Tumor Cells
| Autor: | Mitsuo Umetsu, Yu Katayose, Ryutaro Asano, Hidesuke Fukazawa, Hiroko Kawaguchi, Michiaki Unno, Toshio Kudo, Izumi Kumagai, Takeshi Nakanishi, Hiroki Hayashi, Yasuhiro Watanabe |
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| Rok vydání: | 2007 |
| Předmět: |
CD3 Complex
medicine.medical_treatment CD3 chemical and pharmacologic phenomena CHO Cells Models Biological Biochemistry Immunoglobulin G chemistry.chemical_compound Cricetulus Antigen Cancer immunotherapy Cricetinae Neoplasms Antibodies Bispecific medicine Animals Humans Lymphocytes Cytotoxicity Molecular Biology Cell Proliferation biology Cell Biology Immunotherapy respiratory system Molecular biology ErbB Receptors chemistry Leukocytes Mononuclear biology.protein Growth inhibition Antibody |
| Zdroj: | Journal of Biological Chemistry. 282:27659-27665 |
| ISSN: | 0021-9258 |
| Popis: | We previously reported the marked in vitro and in vivo antitumor activity of hEx3, a humanized diabody (small recombinant bispecific antibody) with epidermal growth factor receptor (EGFR) and CD3 retargeting. Here, we fabricated a tetravalent IgG-like bispecific antibody with two kinds of single-chain Fv (scFv), i.e. humanized anti-EGFR scFv and anti-CD3 scFv, that contains the same four variable domains as hEx3, on the platform of human IgG1 (hEx3-scFv-Fc). hEx3-scFv-Fc prepared from mammalian cells showed specific binding to both EGFR and CD3 target antigens. At one-thousandth (0.1-100 fmol/ml) of the dose of normal hEx3, hEx3-scFv-Fc showed intense cytotoxicity to an EGFR-positive cell line in a growth-inhibition assay using lymphokine-activated killer cells with the T-cell phenotype (T-LAK cells). The enhanced antitumor effect was more clearly observed when peripheral blood mononuclear cells (PBMCs) were used as effector cells, indicating the utility of IgG-like fabrication. These results suggested that the intense antitumor activity is attributable to the multivalency and the presence of the fused human Fc, a hypothesis that was supported by the results of flow cytometry, PBMC proliferation assay, and protein kinase inhibition assay. Furthermore, the growth inhibition effects of hEx3-scFv-Fc were considerably superior to those of the approved therapeutic antibody, cetuximab, which recognizes the same EGFR antigen even when using PBMCs as effector cells. The high potency of hEx3-scFv-Fc may translate into improved antitumor therapy and lower costs of production because of the smaller doses needed. |
| Databáze: | OpenAIRE |
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