title: Development of an anti-claudin-3 and -4 bispecific monoclonal antibody for cancer diagnosis and therapy.
abstract: Most malignant tumors are derived from epithelium, and claudin (CLDN)-3 and CLDN-4 are frequently overexpressed in such tumors. Although antibodies have potential in cancer diagnostics and therapy, development of antibodies against CLDNs has been difficult because the extracellular domains of CLDNs are too small and there is high homology among human, rat, and mouse sequences. Here, we created a monoclonal antibody that recognizes human CLDN-3 and CLDN-4 by immunizing rats with a plasmid vector encoding human CLDN-4. A hybridoma clone that produced a rat monoclonal antibody recognizing both CLDN-3 and -4 (clone 5A5) was obtained from a hybridoma screen by using CLDN-3- and -4-expressing cells; 5A5 did not bind to CLDN-1-, -2-, -5-, -6-, -7-, or -9-expressing cells. Fluorescence-conjugated 5A5 injected into xenograft mice bearing human cancer MKN74 or LoVo cells could visualize the tumor cells. The human-rat chimeric IgG1 monoclonal antibody (xi5A5) activated FcγRIIIa in the presence of CLDN-3- or -4-expressing cells, indicating that xi5A5 may exert antibody-dependent cellular cytotoxicity. Administration of xi5A5 attenuated tumor growth in xenograft mice bearing MKN74 or LoVo cells. These results suggest that 5A5 shows promise in the development of a diagnostic and therapeutic antibody for cancers.
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Xiangru
authors.initials: X
authors.lastname: Li
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Manami
authors.initials: M
authors.lastname: Iida
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Minoru
authors.initials: M
authors.lastname: Tada
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Akihiro
authors.initials: A
authors.lastname: Watari
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Yumi
authors.initials: Y
authors.lastname: Kawahigashi
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Yuka
authors.initials: Y
authors.lastname: Kimura
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Taku
authors.initials: T
authors.lastname: Yamashita
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Akiko
authors.initials: A
authors.lastname: Ishii-Watabe
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Uno
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Masayoshi
authors.initials: M
authors.lastname: Fukasawa
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Hiroki
authors.initials: H
authors.lastname: Kuniyasu
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.).
authors.forename: Kiyohito
authors.initials: K
authors.lastname: Yagi
authors.affiliation: Laboratories of Bio-Functional Molecular Chemistry (X.L., M.I., A.W., Y.Ka., Y.Ki., K.Y., M.K.) and Analytical Chemistry (T.Y., T.U.), Graduate School of Pharmaceutical Sciences, Osaka University, Osaka, Japan; Division of Biological Chemistry and Biologicals, National Institutes of Health Sciences, Tokyo, Japan (M.T., A.I.-W.); Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Tokyo, Japan (M.F.); and Department of Molecular Pathology, Nara Medical University, Nara, Japan (H.K.) masuo@phs.osaka-u.ac.jp.
authors.forename: Masuo
authors.initials: M
authors.lastname: Kondoh
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000627:therapeutic use; D000920:Antibody-Dependent Cell Cytotoxicity; D002460:Cell Line; D045744:Cell Line, Tumor; D062465:Claudin-3 / Q000276:immunology*; D062506:Claudin-4 / Q000276:immunology*; D005260:Female; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology / Q000627:therapeutic use; D007167:Immunotherapy; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009374:Neoplasms, Experimental / Q000175:diagnosis / Q000628:therapy*; D051381:Rats; D017208:Rats, Wistar; D017452:Receptors, IgG / Q000276:immunology; D023041:Xenograft Model Antitumor Assays
chemical_list: D000911:Antibodies, Monoclonal; C568122:CLDN3 protein, human; C568157:CLDN4 protein, human; D062465:Claudin-3; D062506:Claudin-4; D007074:Immunoglobulin G; D017452:Receptors, IgG