Development of an anti-human EphA2 monoclonal antibody Ea2Mab-7 for multiple applications

IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry and Biophysics Reports Pub Date : 2025-04-01 DOI:10.1016/j.bbrep.2025.101998

Hiroyuki Satofuka, Hiroyuki Suzuki, Tomohiro Tanaka, Guanjie Li, Mika K. Kaneko, Yukinari Kato

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引用次数: 0

Abstract

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.

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多用途抗人EphA2单克隆抗体Ea2Mab-7的研制

Ephrin A型受体2 （EphA2）与膜结合配体Ephrin A1、A2和A5结合，引发双向信号。这种信号调节许多生理过程，如组织发育、体内平衡和再生。EphA2-ephrins轴的失调导致多种疾病，包括癌症。EphA2在多种癌症中高表达，促进癌症恶性，而在大多数正常成人组织中其水平相对较低。因此，EphA2是一个很有前景的癌症治疗靶点。本研究采用细胞免疫筛选方法制备了抗人EphA2单克隆抗体。其中，克隆Ea2Mab-7 （IgG1, κ）在流式细胞术中表现出高亲和力和敏感性。测定Ea2Mab-7对CHO/EphA2和MDA-MB-231细胞的解离常数分别为6.2±1.3 × 10−9 M和1.6±0.4 × 10−9 M。Ea2Mab-7可以通过Western blot和免疫组化检测内源性EphA2。因此，Ea2Mab-7在基础研究中具有很强的通用性，有望在抗体治疗和肿瘤诊断等临床应用中有所贡献。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

191

审稿时长

59 days

期刊介绍： Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.