Abstract
We applied the immunoglobulin E (IgE) heavy-chain domain 2 (EHD2) as the covalently linked homodimerization module to generate antibody-scTRAIL fusion proteins. By fusing a humanized single-chain fragment variable (scFv) directed against EGFR to the N-terminus of the EHD2 and a single-chain derivative of TRAIL (scTRAIL) to the C-terminus of the EHD2, we produced a dimeric, tetravalent fusion protein. The fusion protein retained its binding activity for EGFR and TRAIL receptors. In vitro, the targeted antibody-scTRAIL fusion protein exhibited an approximately 8- to 18-fold increased cytotoxic activity compared with the untargeted EHD2-scTRAIL fusion protein. This resulted in increased antitumor activity in a subcutaneous Colo205 xenograft tumor murine model. In summary, the scFv-EHD2-scTRAIL fusion protein combines target cell selectivity with an increased TRAIL activity leading to improved antitumor activities.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Monoclonal, Humanized / pharmacology*
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Apoptosis / genetics
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Apoptosis / immunology
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Cell Line, Tumor
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ErbB Receptors / genetics
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ErbB Receptors / immunology
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Humans
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Immunoglobulin E / genetics
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Immunoglobulin E / immunology
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Immunoglobulin E / pharmacology*
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Mice
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Neoplasms / genetics
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Neoplasms / immunology*
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Neoplasms / therapy
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Receptors, TNF-Related Apoptosis-Inducing Ligand / genetics
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Receptors, TNF-Related Apoptosis-Inducing Ligand / immunology
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / pharmacology
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Single-Chain Antibodies / genetics
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Single-Chain Antibodies / immunology
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Single-Chain Antibodies / pharmacology*
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TNF-Related Apoptosis-Inducing Ligand / genetics*
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TNF-Related Apoptosis-Inducing Ligand / immunology
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Xenograft Model Antitumor Assays
Substances
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Antibodies, Monoclonal, Humanized
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Receptors, TNF-Related Apoptosis-Inducing Ligand
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Recombinant Fusion Proteins
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Single-Chain Antibodies
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TNF-Related Apoptosis-Inducing Ligand
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Immunoglobulin E
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EGFR protein, human
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ErbB Receptors