title: Highly enhanced cytotoxicity of a dimeric bispecific diabody, the hEx3 tetrabody.
abstract: We previously reported the utility for cancer immunotherapy of a humanized bispecific diabody (hEx3) that targets epidermal growth factor receptor and CD3. Here, we used dynamic and static light scattering measurements to show that the multimer fraction observed in hEx3 in solution is a monodisperse tetramer. The multimerization into tetramers increased the inhibition of cancer cell growth by the hEx3 diabody. Furthermore, 1:2 stoichiometric binding for both antigens was observed in a thermodynamic analysis, indicating that the tetramer has bivalent binding activity for each target, and the structure may be in a circular configuration, as is the case for the single-chain Fv tetrabody. In addition to enhanced cytotoxicity, the functional affinity and stability of the hEx3 tetrabody were superior to those of the hEx3 diabody. The increase in molecular weight is also expected to improve the pharmacokinetics of the bispecific diabody, making the hEx3 tetrabody attractive as a therapeutic antibody fragment for cancer immunotherapy.
authors.affiliation: Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, Sendai 980-8579, Japan.
authors.forename: Ryutaro
authors.initials: R
authors.lastname: Asano
authors.forename: Keiko
authors.initials: K
authors.lastname: Ikoma
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Sone
authors.forename: Hiroko
authors.initials: H
authors.lastname: Kawaguchi
authors.forename: Shintaro
authors.initials: S
authors.lastname: Taki
authors.forename: Hiroki
authors.initials: H
authors.lastname: Hayashi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nakanishi
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Umetsu
authors.forename: Yu
authors.initials: Y
authors.lastname: Katayose
authors.forename: Michiaki
authors.initials: M
authors.lastname: Unno
authors.forename: Toshio
authors.initials: T
authors.lastname: Kudo
authors.forename: Izumi
authors.initials: I
authors.lastname: Kumagai
mesh_terms: D018033:Antibodies, Bispecific / Q000235:genetics / Q000627:therapeutic use / Q000633:toxicity*; D000941:Antigens / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology; D002455:Cell Division / Q000187:drug effects; D002850:Chromatography, Gel; D019281:Dimerization; D066246:ErbB Receptors / Q000276:immunology; D006801:Humans; D015979:Killer Cells, Lymphokine-Activated / Q000187:drug effects / Q000276:immunology; D007700:Kinetics; D008027:Light; D009369:Neoplasms / Q000188:drug therapy / Q000276:immunology / Q000473:pathology; D017510:Protein Folding; D011994:Recombinant Proteins / Q000627:therapeutic use; D012542:Scattering, Radiation; D012984:Software; D020349:Surface Plasmon Resonance; D013816:Thermodynamics
chemical_list: D018033:Antibodies, Bispecific; D000941:Antigens; D017252:CD3 Complex; D011994:Recombinant Proteins; D066246:ErbB Receptors
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