Establishment of Novel Anti-EphB3 Monoclonal Antibodies for Multiple Applications
Ephrin type-B receptor 3 (EphB3) binds to transmembrane ephrin-B ligands to regulate cell migration, 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 antihuman EphB3, Eb<sub>3</sub>Mab-5 (IgG<sub>1</sub>, κ) and Eb<sub>3</sub>Mab-11 (IgG<sub>1</sub>, κ), using the Cell-Based Immunization and Screening (CBIS) method. Eb<sub>3</sub>Mab-5 and Eb<sub>3</sub>Mab-11 reacted with EphB3-overexpressed Chinese hamster ovary-K1 (CHO/EphB3) and endogenous EphB3-positive colorectal cancer LS174T in flow cytometry. The apparent binding affinity of Eb<sub>3</sub>Mab-5 for CHO/EphB3 and LS174T was 7.6 × 10<sup>-9</sup> M and 1.7 × 10<sup>-8</sup> M, respectively. Eb<sub>3</sub>Mab-11 could detect EphB3 in western blot analysis and immunohistochemistry. Eb<sub>3</sub>Mab-5 and Eb<sub>3</sub>Mab-11, established by the CBIS method, may contribute to the diagnosis and therapy of EphB3-positive tumors.
Authors
- Mika K. Kaneko (ORCID: https://orcid.org/0000-0002-4158-9208)
- Yukinari Kato (ORCID: https://orcid.org/0000-0001-5385-8201)
- H. Suzuki
- Guanjie Li
Institutions
- Tohoku University (JP)
Publication Details
- Journal
- Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
- Published
- 2025-12-30
- DOI
- https://doi.org/10.1177/21679436251408319
- Primary Topic
- Axon Guidance and Neuronal Signaling
- Type
- article
- Field-Weighted Citation Impact
- 0.00