上皮细胞粘附分子
表位
单克隆抗体
分子生物学
流式细胞术
丙氨酸扫描
丙氨酸
化学
抗体
生物
突变体
生物化学
细胞
氨基酸
免疫学
突变
基因
作者
Guanjie Li,Hiroyuki Suzuki,Tomohiro Tanaka,Teizo Asano,Takeo Yoshikawa,Mika K. Kaneko,Yukinari Kato
出处
期刊:Monoclonal antibodies in immunodiagnosis and immunotherapy
[Mary Ann Liebert]
日期:2023-02-01
卷期号:42 (1): 41-47
被引量:2
标识
DOI:10.1089/mab.2022.0031
摘要
The epithelial cell adhesion molecule (EpCAM) is a type I transmembrane glycoprotein, and plays critical roles in cell adhesion, proliferation, and tumorigenesis. EpCAM has been considered as a promising target for tumor diagnosis and therapy. Anti-EpCAM monoclonal antibodies (mAbs) have been developed for EpCAM-overexpressed tumors, and several clinical trials have demonstrated promising outcomes. We previously established an anti-EpCAM mAb, EpMab-37 (mouse IgG1, kappa), using the Cell-Based Immunization and Screening method. EpMab-37 was revealed to recognize the conformational epitope of EpCAM. In this study, we determined the critical epitope of EpMab-37 by flow cytometry using the 1 × alanine scanning (1 × Ala-scan) and the 2 × alanine scanning (2 × Ala-scan) method. We first performed flow cytometry by 1 × Ala-scan using one alanine (or glycine)-substituted EpCAM mutants, which were expressed on Chinese hamster ovary-K1 cells, and found that the EpMab-37 did not recognize the R163A mutant of EpCAM. We next performed flow cytometry by 2 × Ala-scan using two alanine (or glycine) residues-substituted EpCAM mutants, and confirmed that EpMab-37 did not recognize R163A-including mutants of EpCAM. The results indicated that the critical binding epitope of EpMab-37 includes Arg163 of EpCAM.
科研通智能强力驱动
Strongly Powered by AbleSci AI