{"title":"新型抗人CCR9单克隆抗体(C9Mab-11)的2 ×丙氨酸扫描表位定位","authors":"Yu Isoda, Tomohiro Tanaka, Hiroyuki Suzuki, Teizo Asano, Kaishi Kitamura, Yuma Kudo, Ryo Ejima, Kazuki Ozawa, Takeo Yoshikawa, Mika K Kaneko, Yukinari Kato","doi":"10.1089/mab.2022.0035","DOIUrl":null,"url":null,"abstract":"<p><p>We recently developed a novel anti-human C-C chemokine receptor 9 (hCCR9) monoclonal antibody (mAb), C<sub>9</sub>Mab-11, which is applicable to flow cytometry, western blotting, and enzyme-linked immunosorbent assay (ELISA). This study aims to identify the binding epitope of C<sub>9</sub>Mab-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 C<sub>9</sub>Mab-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 C<sub>9</sub>Mab-11 includes Pro11, Asn12, Met13, and Ala14 of hCCR9, with the region around Met13 being particularly important. The successful identification of the C<sub>9</sub>Mab-11 epitope might be useful for the future pathophysiological analysis of hCCR9.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":"42 2","pages":"73-76"},"PeriodicalIF":0.0000,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Epitope Mapping of the Novel Anti-Human CCR9 Monoclonal Antibody (C<sub>9</sub>Mab-11) by 2 × Alanine Scanning.\",\"authors\":\"Yu Isoda, Tomohiro Tanaka, Hiroyuki Suzuki, Teizo Asano, Kaishi Kitamura, Yuma Kudo, Ryo Ejima, Kazuki Ozawa, Takeo Yoshikawa, Mika K Kaneko, Yukinari Kato\",\"doi\":\"10.1089/mab.2022.0035\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>We recently developed a novel anti-human C-C chemokine receptor 9 (hCCR9) monoclonal antibody (mAb), C<sub>9</sub>Mab-11, which is applicable to flow cytometry, western blotting, and enzyme-linked immunosorbent assay (ELISA). This study aims to identify the binding epitope of C<sub>9</sub>Mab-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 C<sub>9</sub>Mab-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 C<sub>9</sub>Mab-11 includes Pro11, Asn12, Met13, and Ala14 of hCCR9, with the region around Met13 being particularly important. The successful identification of the C<sub>9</sub>Mab-11 epitope might be useful for the future pathophysiological analysis of hCCR9.</p>\",\"PeriodicalId\":53514,\"journal\":{\"name\":\"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy\",\"volume\":\"42 2\",\"pages\":\"73-76\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1089/mab.2022.0035\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"Medicine\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1089/mab.2022.0035","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}
Epitope Mapping of the Novel Anti-Human CCR9 Monoclonal Antibody (C9Mab-11) by 2 × Alanine Scanning.
We recently developed a novel anti-human C-C chemokine receptor 9 (hCCR9) monoclonal antibody (mAb), C9Mab-11, which is applicable to flow cytometry, western blotting, and enzyme-linked immunosorbent assay (ELISA). This study aims to identify the binding epitope of C9Mab-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 C9Mab-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 C9Mab-11 includes Pro11, Asn12, Met13, and Ala14 of hCCR9, with the region around Met13 being particularly important. The successful identification of the C9Mab-11 epitope might be useful for the future pathophysiological analysis of hCCR9.
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