Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, Sendai, Japan.
Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Monoclon Antib Immunodiagn Immunother. 2022 Feb;41(1):20-26. doi: 10.1089/mab.2021.0051.
CD20, which is expressed on B lymphocytes, has been studied as a therapeutic target for B cell lymphomas and autoimmune disorders. Identifying the binding epitopes of monoclonal antibodies (mAbs) can contribute to our understanding of their functions. We have previously developed an anti-CD20 mAb (clone CMab-11) using a Cell-Based Immunization and Screening (CBIS) method. In this study, we aimed to determine the binding epitopes of anti-CD20 mAbs, such as CMab-11 and 2H7, using the His-tag insertion for epitope mapping (HisMAP). The results showed that -NPSE- and -EPANPSE- in the second loop of CD20 were essential for CMab-11 binding and 2H7 binding, respectively. Although we developed many mAbs that recognize conformational epitopes using the CBIS method, there are many difficulties in epitope mapping for these mAbs. HisMAP could be useful for determining the conformational epitopes of other mAbs against membrane proteins.
CD20 表达于 B 淋巴细胞上，已被研究作为治疗 B 细胞淋巴瘤和自身免疫性疾病的靶点。鉴定单克隆抗体（mAb）的结合表位有助于我们了解其功能。我们先前使用基于细胞的免疫接种和筛选（CBIS）方法开发了一种抗 CD20 mAb（克隆 CMab-11）。在这项研究中，我们旨在使用 His 标签插入进行表位作图（HisMAP）来确定抗 CD20 mAb，如 CMab-11 和 2H7 的结合表位。结果表明，CD20 第二环中的 -NPSE- 和 -EPANPSE- 对于 CMab-11 结合和 2H7 结合分别是必需的。虽然我们使用 CBIS 方法开发了许多识别构象表位的 mAb，但对于这些 mAb 进行表位作图存在许多困难。HisMAP 可用于确定针对膜蛋白的其他 mAb 的构象表位。
