Development of a Novel Anti-human EphA1 Monoclonal Antibody, Ea1Mab-30, for Multiple Applications.

Q3 Medicine
Tomohiro Tanaka, Hiroyuki Suzuki, Honoka Taruta, Ayano Saga, Guanjie Li, Shiori Fujisawa, Mika K Kaneko, Yukinari Kato
{"title":"Development of a Novel Anti-human EphA1 Monoclonal Antibody, Ea<sub>1</sub>Mab-30, for Multiple Applications.","authors":"Tomohiro Tanaka, Hiroyuki Suzuki, Honoka Taruta, Ayano Saga, Guanjie Li, Shiori Fujisawa, Mika K Kaneko, Yukinari Kato","doi":"10.1089/mab.2025.0006","DOIUrl":null,"url":null,"abstract":"<p><p>Erythropoietin-producing hepatocellular receptor A1 (EphA1) is one of the Eph receptor family members, the largest group of receptor tyrosine kinases. EphA1 is expressed in various tissues and regulates cellular homeostasis by interacting with its membrane-bound ephrin ligands and other receptors. EphA1 critically correlates with the pathogenesis in several disorders, including Alzheimer's disease and cancers. Therefore, establishing sensitive monoclonal antibodies (mAbs) for EphA1 has been desired for basic research, diagnosis, and treatment. In this study, a novel specific and sensitive anti-human EphA1 mAb, clone Ea<sub>1</sub>Mab-30 (mouse IgG<sub>1</sub>, kappa), was established by the Cell-Based Immunization and Screening (CBIS) method. Ea<sub>1</sub>Mab-30 demonstrated reactivity with an EphA1-overexpressed Chinese hamster ovary-K1 cell line (CHO/EphA1), an endogenously EphA1-expressing bladder carcinoma cell line (5637), and a colorectal adenocarcinoma cell line (Caco-2) in flow cytometry. Crossreactivities of Ea<sub>1</sub>Mab-30 with other Eph receptors were not observed. Furthermore, the values of apparent binding affinity for CHO/EphA1 and 5637 were determined to be 8.9 × 10<sup>-9</sup> M and 1.7 × 10<sup>-9</sup> M, respectively. Furthermore, Ea<sub>1</sub>Mab-30 detected EphA1 protein in CHO/EphA1 and 5637 lysates using Western blot analysis. Ea<sub>1</sub>Mab-30 also clearly stained EphA1 of formalin-fixed paraffin-embedded CHO/EphA1 using immunohistochemistry. Ea<sub>1</sub>Mab-30, established by CBIS method, could help analyze the EphA1-contributed cellular functions and have potential applications in pathological diagnosis and treatment with specificity and high affinity for EphA1-expressing cells.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":"44 3","pages":"41-52"},"PeriodicalIF":0.0000,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1089/mab.2025.0006","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 0

Abstract

Erythropoietin-producing hepatocellular receptor A1 (EphA1) is one of the Eph receptor family members, the largest group of receptor tyrosine kinases. EphA1 is expressed in various tissues and regulates cellular homeostasis by interacting with its membrane-bound ephrin ligands and other receptors. EphA1 critically correlates with the pathogenesis in several disorders, including Alzheimer's disease and cancers. Therefore, establishing sensitive monoclonal antibodies (mAbs) for EphA1 has been desired for basic research, diagnosis, and treatment. In this study, a novel specific and sensitive anti-human EphA1 mAb, clone Ea1Mab-30 (mouse IgG1, kappa), was established by the Cell-Based Immunization and Screening (CBIS) method. Ea1Mab-30 demonstrated reactivity with an EphA1-overexpressed Chinese hamster ovary-K1 cell line (CHO/EphA1), an endogenously EphA1-expressing bladder carcinoma cell line (5637), and a colorectal adenocarcinoma cell line (Caco-2) in flow cytometry. Crossreactivities of Ea1Mab-30 with other Eph receptors were not observed. Furthermore, the values of apparent binding affinity for CHO/EphA1 and 5637 were determined to be 8.9 × 10-9 M and 1.7 × 10-9 M, respectively. Furthermore, Ea1Mab-30 detected EphA1 protein in CHO/EphA1 and 5637 lysates using Western blot analysis. Ea1Mab-30 also clearly stained EphA1 of formalin-fixed paraffin-embedded CHO/EphA1 using immunohistochemistry. Ea1Mab-30, established by CBIS method, could help analyze the EphA1-contributed cellular functions and have potential applications in pathological diagnosis and treatment with specificity and high affinity for EphA1-expressing cells.

多用途抗人EphA1单克隆抗体Ea1Mab-30的研制
促红细胞生成素产生肝细胞受体A1 (EphA1)是Eph受体家族成员之一,是酪氨酸激酶受体中最大的一类。EphA1在多种组织中表达,并通过与其膜结合的ephrin配体和其他受体相互作用来调节细胞稳态。EphA1与多种疾病的发病机制密切相关,包括阿尔茨海默病和癌症。因此,建立敏感的EphA1单克隆抗体(mab)已成为基础研究、诊断和治疗的需要。本研究通过细胞免疫筛选(Cell-Based Immunization and Screening, CBIS)方法,建立了一种新的特异性、敏感性的抗人EphA1单抗克隆Ea1Mab-30(小鼠IgG1, kappa)。流式细胞术显示Ea1Mab-30对EphA1过表达的中国仓鼠卵巢k1细胞系(CHO/EphA1)、内源性表达EphA1的膀胱癌细胞系(5637)和结直肠腺癌细胞系(Caco-2)具有反应性。Ea1Mab-30未与其他Eph受体发生交叉反应。此外,对CHO/EphA1和5637的表观结合亲和力分别为8.9 × 10-9 M和1.7 × 10-9 M。此外,Ea1Mab-30采用Western blot方法检测CHO/EphA1和5637裂解物中的EphA1蛋白。Ea1Mab-30也通过免疫组化对福尔马林固定石蜡包埋CHO/EphA1的EphA1进行了清晰的染色。通过CBIS方法建立的Ea1Mab-30对表达epha1的细胞具有特异性和高亲和力,有助于分析epha1介导的细胞功能,在病理诊断和治疗方面具有潜在的应用前景。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
CiteScore
4.80
自引率
0.00%
发文量
49
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信