Hiroyuki Suzuki, Tomohiro Tanaka, Guanjie Li, Tsunenori Ouchida, Mika K Kaneko, Yukinari Kato
{"title":"为流式细胞仪开发灵敏的抗小鼠 CCR5 单克隆抗体。","authors":"Hiroyuki Suzuki, Tomohiro Tanaka, Guanjie Li, Tsunenori Ouchida, Mika K Kaneko, Yukinari Kato","doi":"10.1089/mab.2024.0004","DOIUrl":null,"url":null,"abstract":"<p><p>C-C chemokine receptor 5 (CCR5), a member of the G protein-coupled receptor family, is the most common coreceptor for the human immunodeficiency virus type 1. CCR5 is also involved in the pathogenesis of tumors and inflammatory diseases. The CCR5 antagonists including monoclonal antibodies (mAbs) have been developed and evaluated in clinical trials. In this study, we developed novel mAbs for mouse CCR5 (mCCR5) using the Cell-Based Immunization and Screening (CBIS) method. One of the established anti-mCCR5 mAbs, C<sub>5</sub>Mab-2 (rat IgG<sub>2b</sub>, kappa), reacted with mCCR5-overexpressed Chinese hamster ovary-K1 (CHO/mCCR5) and an endogenously mCCR5-expressing cell line (L1210) by flow cytometry. Using flow cytometry, the dissociation constant (<i>K</i><sub>D</sub>) of C<sub>5</sub>Mab-2 for CHO/mCCR5 was determined as 4.3 × 10<sup>-8</sup> M. These results indicated that C<sub>5</sub>Mab-2 is useful for the detection of mCCR5 in flow cytometry and may be applicable to obtain the proof of concept in preclinical studies.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Development of a Sensitive Anti-Mouse CCR5 Monoclonal Antibody for Flow Cytometry.\",\"authors\":\"Hiroyuki Suzuki, Tomohiro Tanaka, Guanjie Li, Tsunenori Ouchida, Mika K Kaneko, Yukinari Kato\",\"doi\":\"10.1089/mab.2024.0004\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>C-C chemokine receptor 5 (CCR5), a member of the G protein-coupled receptor family, is the most common coreceptor for the human immunodeficiency virus type 1. CCR5 is also involved in the pathogenesis of tumors and inflammatory diseases. The CCR5 antagonists including monoclonal antibodies (mAbs) have been developed and evaluated in clinical trials. In this study, we developed novel mAbs for mouse CCR5 (mCCR5) using the Cell-Based Immunization and Screening (CBIS) method. One of the established anti-mCCR5 mAbs, C<sub>5</sub>Mab-2 (rat IgG<sub>2b</sub>, kappa), reacted with mCCR5-overexpressed Chinese hamster ovary-K1 (CHO/mCCR5) and an endogenously mCCR5-expressing cell line (L1210) by flow cytometry. Using flow cytometry, the dissociation constant (<i>K</i><sub>D</sub>) of C<sub>5</sub>Mab-2 for CHO/mCCR5 was determined as 4.3 × 10<sup>-8</sup> M. These results indicated that C<sub>5</sub>Mab-2 is useful for the detection of mCCR5 in flow cytometry and may be applicable to obtain the proof of concept in preclinical studies.</p>\",\"PeriodicalId\":53514,\"journal\":{\"name\":\"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-08-01\",\"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.2024.0004\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2024/6/5 0:00:00\",\"PubModel\":\"Epub\",\"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.2024.0004","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/6/5 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}
Development of a Sensitive Anti-Mouse CCR5 Monoclonal Antibody for Flow Cytometry.
C-C chemokine receptor 5 (CCR5), a member of the G protein-coupled receptor family, is the most common coreceptor for the human immunodeficiency virus type 1. CCR5 is also involved in the pathogenesis of tumors and inflammatory diseases. The CCR5 antagonists including monoclonal antibodies (mAbs) have been developed and evaluated in clinical trials. In this study, we developed novel mAbs for mouse CCR5 (mCCR5) using the Cell-Based Immunization and Screening (CBIS) method. One of the established anti-mCCR5 mAbs, C5Mab-2 (rat IgG2b, kappa), reacted with mCCR5-overexpressed Chinese hamster ovary-K1 (CHO/mCCR5) and an endogenously mCCR5-expressing cell line (L1210) by flow cytometry. Using flow cytometry, the dissociation constant (KD) of C5Mab-2 for CHO/mCCR5 was determined as 4.3 × 10-8 M. These results indicated that C5Mab-2 is useful for the detection of mCCR5 in flow cytometry and may be applicable to obtain the proof of concept in preclinical studies.
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