Establishment of Novel Anti-EphB3 Monoclonal Antibodies for Multiple Applications
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Ephrin type-B receptor 3 (EphB3) binds to transmembrane ephrin-B ligands to regulate cell mi-gration, adhesion, and proliferation. EphB3 exhibits a gradient expression pattern in the normal intestine, with the highest levels at the crypt base, and plays a crucial role in the maintenance of normal intestinal epithelium. Therefore, anti-EphB3 monoclonal antibodies (mAbs) are required for basic research and diagnosis. In this study, we developed novel anti-human EphB3, Eb3Mab-5 (IgG1, κ) and Eb3Mab-11 (IgG1, κ), using the Cell-Based Immunization and Screening (CBIS) method. Eb3Mab-5 and Eb3Mab-11 reacted with EphB3-overexpressed Chinese hamster ova-ry-K1 (CHO/EphB3) and endogenous EphB3-positive colorectal cancer LS174T in flow cytometry. The KD values of Eb3Mab-5 for CHO/EphB3 and LS174T were 7.6 ×10-9 M and 1.7 ×10-8 M, re-spectively. Eb3Mab-11 could detect EphB3 in western blot analysis and immunohistochemistry. Eb3Mab-5 and Eb3Mab-11 may contribute to the diagnosis and therapy of EphB3-positive tu-mors.