Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Sendai, Japan.
Department of Molecular Pharmacology, and Tohoku University Graduate School of Medicine, Sendai, Japan.
Monoclon Antib Immunodiagn Immunother. 2023 Apr;42(2):73-76. doi: 10.1089/mab.2022.0035.
We recently developed a novel anti-human C-C chemokine receptor 9 (hCCR9) monoclonal antibody (mAb), CMab-11, which is applicable to flow cytometry, western blotting, and enzyme-linked immunosorbent assay (ELISA). This study aims to identify the binding epitope of CMab-11 by using 1 × and 2 × alanine (or glycine) substituted-hCCR9 peptides (1 × and 2 × Ala-scan) by ELISA. According to the 1 × Ala-scan analysis, the response of CMab-11 was diminished against M13A of the hCCR9 peptide, but was not eliminated. In the 2 × Ala-scan analysis, the reactions were abolished in the substitution of P11A-N12A, N12A-M13A, and M13A-A14G of hCCR9 N-terminal peptides. The results indicate that the binding epitope of CMab-11 includes Pro11, Asn12, Met13, and Ala14 of hCCR9, with the region around Met13 being particularly important. The successful identification of the CMab-11 epitope might be useful for the future pathophysiological analysis of hCCR9.
我们最近开发了一种新型抗人 C-C 趋化因子受体 9（hCCR9）单克隆抗体（mAb）CMab-11，它适用于流式细胞术、蛋白质印迹和酶联免疫吸附测定（ELISA）。本研究旨在通过 ELISA 用 1×和 2×丙氨酸（或甘氨酸）取代 hCCR9 肽（1×和 2×Ala-scan）来鉴定 CMab-11 的结合表位。根据 1×Ala-scan 分析，CMab-11 对 hCCR9 肽的 M13A 的反应性降低，但未被消除。在 2×Ala-scan 分析中，在 hCCR9 N 末端肽的 P11A-N12A、N12A-M13A 和 M13A-A14G 取代时，反应被消除。结果表明，CMab-11 的结合表位包括 hCCR9 的 Pro11、Asn12、Met13 和 Ala14，其中 Met13 周围的区域尤为重要。CMab-11 表位的成功鉴定可能对 hCCR9 的未来病理生理分析有用。
