Ren Nanamiya, Hiroyuki Suzuki, Mika K. Kaneko, and Yukinari Kato. Development of an Anti-EphB4 Monoclonal Antibody for Multiple Applications Against Breast Cancers. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy.ahead of print http://doi.org/10.1089/mab.2023.0015
Ren Nanamiya, Hiroyuki Suzuki, Mika K. Kaneko, and Yukinari Kato. Development of an Anti-EphB4 Monoclonal Antibody for Multiple Applications Against Breast Cancers. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy.ahead of print http://doi.org/10.1089/mab.2023.0015
Ren Nanamiya, Hiroyuki Suzuki, Mika K. Kaneko, and Yukinari Kato. Development of an Anti-EphB4 Monoclonal Antibody for Multiple Applications Against Breast Cancers. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy.ahead of print http://doi.org/10.1089/mab.2023.0015
Ren Nanamiya, Hiroyuki Suzuki, Mika K. Kaneko, and Yukinari Kato. Development of an Anti-EphB4 Monoclonal Antibody for Multiple Applications Against Breast Cancers. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy.ahead of print http://doi.org/10.1089/mab.2023.0015
Abstract
The erythropoietin-producing hepatocellular carcinoma (Eph) receptors are the largest receptor tyrosine kinases family. EphB4 is essential for cell adhesion and motility during embryogenesis. Pathologically, EphB4 is overexpressed and contributes to poor prognosis in various tumors. Therefore, sensitive monoclonal antibodies (mAbs) should be developed to predict the prognosis for multiple tumors with high EphB4 expression, including breast and gastric cancers. This study aimed to develop highly sensitive and specific anti-EphB4 mAbs for several applications using the Cell-Based Immunization and Screening (CBIS) method. EphB4-overexpressed Chinese hamster ovary (CHO)-K1 (CHO/EphB4) cells were immunized into mice, and we established an anti-EphB4 mAb (clone B4Mab-7), which is applicable for flow cytometry, western blotting, and immunohistochemistry. B4Mab-7 reacted with endogenous EphB4-positive breast cancer cell lines, MCF-7 and MDA-MB-468, but did not react with EphB4-knockout MCF-7 (BINDS-52) in flow cytometry. Dissociation constant (KD) values were determined to be 2.9 × 10‑9 M, 1.3 × 10‑9 M, and 3.3 × 10‑9 M by flow cytometric analysis for CHO/EphB4, MCF-7, and MDA-MB-468 cells, respectively. B4Mab-7 detected the EphB4 protein bands from breast cancer cells in western blotting, and stained breast cancer tissues immunohistochemistry. Altogether, B4Mab-7 demonstrated high sensitivity and specificity against EphB4 in various applications.
Keywords
EphB4; monoclonal antibody; Cell-Based Immunization and Screening; immunohistochemistry
Subject
Medicine and Pharmacology, Oncology and Oncogenics
Copyright:
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