Development of an anti-human EphA2 monoclonal antibody Ea2Mab-7 for multiple applications
Ephrin type A receptor 2 (EphA2) binds to membrane-bound ligands, ephrin A1, A2, and A5, eliciting bidirectional signaling. This signaling regulates many physiological processes, such as tissue development, homeostasis, and regeneration. The dysregulation of the EphA2-ephrins axis contributes to var...
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Elsevier
2025-06-01
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| Series: | Biochemistry and Biophysics Reports |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580825000858 |
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| author | Hiroyuki Satofuka Hiroyuki Suzuki Tomohiro Tanaka Guanjie Li Mika K. Kaneko Yukinari Kato |
| author_facet | Hiroyuki Satofuka Hiroyuki Suzuki Tomohiro Tanaka Guanjie Li Mika K. Kaneko Yukinari Kato |
| author_sort | Hiroyuki Satofuka |
| collection | DOAJ |
| description | Ephrin type A receptor 2 (EphA2) binds to membrane-bound ligands, ephrin A1, A2, and A5, eliciting bidirectional signaling. This signaling regulates many physiological processes, such as tissue development, homeostasis, and regeneration. The dysregulation of the EphA2-ephrins axis contributes to various diseases, including cancers. The high expression of EphA2 is observed in various cancers, which promotes cancer malignancy, whereas its levels are relatively low in most normal adult tissues. Therefore, EphA2 is a promising target for cancer therapy. We developed anti-human EphA2 monoclonal antibodies in this study using the Cell-Based Immunization and Screening method. Among them, a clone Ea2Mab-7 (IgG1, κ) exhibited a high affinity and sensitivity in flow cytometry. The dissociation constant values of Ea2Mab-7 for CHO/EphA2 and MDA-MB-231 cells were determined as 6.2 ± 1.3 × 10−9 M and 1.6 ± 0.4 × 10−9 M, respectively. Furthermore, Ea2Mab-7 can detect endogenous EphA2 in Western blot and immunohistochemistry. Therefore, the Ea2Mab-7 is highly versatile for basic research and is expected to contribute to clinical applications, such as antibody therapy and tumor diagnosis. |
| format | Article |
| id | doaj-art-3a3ddc68f783413aaa7620c7cd452717 |
| institution | OA Journals |
| issn | 2405-5808 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
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| series | Biochemistry and Biophysics Reports |
| spelling | doaj-art-3a3ddc68f783413aaa7620c7cd4527172025-08-20T02:17:13ZengElsevierBiochemistry and Biophysics Reports2405-58082025-06-014210199810.1016/j.bbrep.2025.101998Development of an anti-human EphA2 monoclonal antibody Ea2Mab-7 for multiple applicationsHiroyuki Satofuka0Hiroyuki Suzuki1Tomohiro Tanaka2Guanjie Li3Mika K. Kaneko4Yukinari Kato5Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, JapanDepartment of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, JapanDepartment of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, JapanDepartment of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, JapanDepartment of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, JapanCorresponding author.; Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi, 980-8575, JapanEphrin type A receptor 2 (EphA2) binds to membrane-bound ligands, ephrin A1, A2, and A5, eliciting bidirectional signaling. This signaling regulates many physiological processes, such as tissue development, homeostasis, and regeneration. The dysregulation of the EphA2-ephrins axis contributes to various diseases, including cancers. The high expression of EphA2 is observed in various cancers, which promotes cancer malignancy, whereas its levels are relatively low in most normal adult tissues. Therefore, EphA2 is a promising target for cancer therapy. We developed anti-human EphA2 monoclonal antibodies in this study using the Cell-Based Immunization and Screening method. Among them, a clone Ea2Mab-7 (IgG1, κ) exhibited a high affinity and sensitivity in flow cytometry. The dissociation constant values of Ea2Mab-7 for CHO/EphA2 and MDA-MB-231 cells were determined as 6.2 ± 1.3 × 10−9 M and 1.6 ± 0.4 × 10−9 M, respectively. Furthermore, Ea2Mab-7 can detect endogenous EphA2 in Western blot and immunohistochemistry. Therefore, the Ea2Mab-7 is highly versatile for basic research and is expected to contribute to clinical applications, such as antibody therapy and tumor diagnosis.http://www.sciencedirect.com/science/article/pii/S2405580825000858EphA2Monoclonal antibodyCell-Based Immunization and ScreeningFlow cytometryImmunohistochemistryBreast cancer |
| spellingShingle | Hiroyuki Satofuka Hiroyuki Suzuki Tomohiro Tanaka Guanjie Li Mika K. Kaneko Yukinari Kato Development of an anti-human EphA2 monoclonal antibody Ea2Mab-7 for multiple applications Biochemistry and Biophysics Reports EphA2 Monoclonal antibody Cell-Based Immunization and Screening Flow cytometry Immunohistochemistry Breast cancer |
| title | Development of an anti-human EphA2 monoclonal antibody Ea2Mab-7 for multiple applications |
| title_full | Development of an anti-human EphA2 monoclonal antibody Ea2Mab-7 for multiple applications |
| title_fullStr | Development of an anti-human EphA2 monoclonal antibody Ea2Mab-7 for multiple applications |
| title_full_unstemmed | Development of an anti-human EphA2 monoclonal antibody Ea2Mab-7 for multiple applications |
| title_short | Development of an anti-human EphA2 monoclonal antibody Ea2Mab-7 for multiple applications |
| title_sort | development of an anti human epha2 monoclonal antibody ea2mab 7 for multiple applications |
| topic | EphA2 Monoclonal antibody Cell-Based Immunization and Screening Flow cytometry Immunohistochemistry Breast cancer |
| url | http://www.sciencedirect.com/science/article/pii/S2405580825000858 |
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