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Antibodies 2013, 2(2), 338-352;

Immunotherapy of B-Cell Lymphoma with an Engineered Bispecific Antibody Targeting CD19 and CD5

Department of Translational Immunology, German Cancer Research Center and National Center for Tumor Diseases, 69120 Heidelberg, Germany
Department of Tumor Genetics and Immunogenetics, Max-Delbrück-Center of Molecular Medicine, 13125 Berlin, Germany
Molecular Immunotherapy, Max-Delbrück Center of Molecular Medicine, 13125 Berlin, Germany
Department of Hematology, Oncology and Tumor Immunology, Charité Universitätsmedizin, 13353 Berlin, Germany
Department Peptide Arrays and Antibody Libraries, Karlsruhe Institute of Technology (KIT), 76344 Eggenstein-Leopoldshafen, Germany
Author to whom correspondence should be addressed.
Received: 26 March 2013 / Revised: 22 April 2013 / Accepted: 22 April 2013 / Published: 14 May 2013
(This article belongs to the Special Issue Bispecific Antibodies for Dual Targeting Strategies)
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Using genetic engineering a humanized Fab fragment with specificity for CD19 was fused to a disulfide-stabilized single-chain antibody (dsFv) recognizing CD5. This format should show reduced immunogenicity and improved tissue penetration. The specificity of bsAb FabCD19xdsFvCD5 binding to target cells was verified by flow cytometry on B and T lymphoma cell lines. Binding affinities of both arms were compared with the bivalent parental antibodies against CD19 and CD5 by binding competition assay. Redirected lysis of B lymphoma cells by preactivated PBMC from healthy donors was demonstrated in a chromium-release assay. A clear dose-response relationship could be established in the range from 1 ng/mL to 10 mg/mL bsAb. To evaluate the in vivo efficacy of bsAb FabCD19xdsFvCD5, NOD/SCID mice were intravenously injected with luciferase transfected Raji lymphoma cells together with pre-activated PBMC. Mice received five injections of therapeutic bsAb or control antibodies. While in the control groups all mice died within 40 to 50 days, 40% of bsAb treated animals survived longer than 60 days. View Full-Text
Keywords: bispecific antibody; lymphoma targeting; immunotherapy; CD19; CD5 bispecific antibody; lymphoma targeting; immunotherapy; CD19; CD5

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Lüttgau, S.; Deppe, D.; Meyer, S.; Fertig, R.; Panjideh, H.; Lipp, M.; Schmetzer, O.; Pezzutto, A.; Breitling, F.; Moldenhauer, G. Immunotherapy of B-Cell Lymphoma with an Engineered Bispecific Antibody Targeting CD19 and CD5. Antibodies 2013, 2, 338-352.

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