Miu Hirose, Hiroyuki Suzuki, Rena Ubukata, Tomohiro Tanaka, Mika K. Kaneko, Yukinari Kato
{"title":"开发特异性抗小鼠非典型趋化因子受体 4 单克隆抗体","authors":"Miu Hirose, Hiroyuki Suzuki, Rena Ubukata, Tomohiro Tanaka, Mika K. Kaneko, Yukinari Kato","doi":"10.1016/j.bbrep.2024.101824","DOIUrl":null,"url":null,"abstract":"<div><p>Leukocyte migration is an essential function of innate and adaptive immune responses. Chemokines and their receptors control the migration system. The abundance of chemokines is controlled by atypical chemokine receptors (ACKRs), chemokine receptor-like molecules that do not couple to the G protein signaling pathways. Among them, ACKR4 regulates dendritic cell migration by controlling the ligands and is involved in tumor development in mouse models. Because no anti-mouse ACKR4 (mACKR4) monoclonal antibody (mAb) for flow cytometry has been reported, this study aimed to develop a novel mAb for mACKR4. Among the established anti-mACKR4 mAbs, A<sub>4</sub>Mab-1 (rat IgG<sub>2b</sub>, kappa), A<sub>4</sub>Mab-2 (rat IgG<sub>2b</sub>, kappa), and A<sub>4</sub>Mab-3 (rat IgG<sub>2b</sub>, kappa) recognized mACKR4-overexpressed Chinese hamster ovary-K1 (CHO/mACKR4) by flow cytometry. The dissociation constant (<em>K</em><sub>D</sub>) values of A<sub>4</sub>Mab-1, A<sub>4</sub>Mab-2, and A<sub>4</sub>Mab-3 for CHO/mACKR4 were determined as 6.0 × 10<sup>−9</sup> M, 1.3 × 10<sup>−8</sup> M, and 1.7 × 10<sup>−9</sup> M, respectively. Furthermore, A<sub>4</sub>Mab-1 and A<sub>4</sub>Mab-2 could detect mACKR4 by western blotting. These results indicated that A<sub>4</sub>Mab-1, A<sub>4</sub>Mab-2, and A<sub>4</sub>Mab-3 help to detect mACKR4 by flow cytometry and western blotting and obtain the proof of concept in preclinical models.</p></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"40 ","pages":"Article 101824"},"PeriodicalIF":2.3000,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2405580824001882/pdfft?md5=df1bec1ed4b9eaea11a06516e0feac67&pid=1-s2.0-S2405580824001882-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Development of specific anti-mouse atypical chemokine receptor 4 monoclonal antibodies\",\"authors\":\"Miu Hirose, Hiroyuki Suzuki, Rena Ubukata, Tomohiro Tanaka, Mika K. Kaneko, Yukinari Kato\",\"doi\":\"10.1016/j.bbrep.2024.101824\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Leukocyte migration is an essential function of innate and adaptive immune responses. Chemokines and their receptors control the migration system. The abundance of chemokines is controlled by atypical chemokine receptors (ACKRs), chemokine receptor-like molecules that do not couple to the G protein signaling pathways. Among them, ACKR4 regulates dendritic cell migration by controlling the ligands and is involved in tumor development in mouse models. Because no anti-mouse ACKR4 (mACKR4) monoclonal antibody (mAb) for flow cytometry has been reported, this study aimed to develop a novel mAb for mACKR4. Among the established anti-mACKR4 mAbs, A<sub>4</sub>Mab-1 (rat IgG<sub>2b</sub>, kappa), A<sub>4</sub>Mab-2 (rat IgG<sub>2b</sub>, kappa), and A<sub>4</sub>Mab-3 (rat IgG<sub>2b</sub>, kappa) recognized mACKR4-overexpressed Chinese hamster ovary-K1 (CHO/mACKR4) by flow cytometry. The dissociation constant (<em>K</em><sub>D</sub>) values of A<sub>4</sub>Mab-1, A<sub>4</sub>Mab-2, and A<sub>4</sub>Mab-3 for CHO/mACKR4 were determined as 6.0 × 10<sup>−9</sup> M, 1.3 × 10<sup>−8</sup> M, and 1.7 × 10<sup>−9</sup> M, respectively. Furthermore, A<sub>4</sub>Mab-1 and A<sub>4</sub>Mab-2 could detect mACKR4 by western blotting. These results indicated that A<sub>4</sub>Mab-1, A<sub>4</sub>Mab-2, and A<sub>4</sub>Mab-3 help to detect mACKR4 by flow cytometry and western blotting and obtain the proof of concept in preclinical models.</p></div>\",\"PeriodicalId\":8771,\"journal\":{\"name\":\"Biochemistry and Biophysics Reports\",\"volume\":\"40 \",\"pages\":\"Article 101824\"},\"PeriodicalIF\":2.3000,\"publicationDate\":\"2024-09-07\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2405580824001882/pdfft?md5=df1bec1ed4b9eaea11a06516e0feac67&pid=1-s2.0-S2405580824001882-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biochemistry and Biophysics Reports\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2405580824001882\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochemistry and Biophysics Reports","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2405580824001882","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Development of specific anti-mouse atypical chemokine receptor 4 monoclonal antibodies
Leukocyte migration is an essential function of innate and adaptive immune responses. Chemokines and their receptors control the migration system. The abundance of chemokines is controlled by atypical chemokine receptors (ACKRs), chemokine receptor-like molecules that do not couple to the G protein signaling pathways. Among them, ACKR4 regulates dendritic cell migration by controlling the ligands and is involved in tumor development in mouse models. Because no anti-mouse ACKR4 (mACKR4) monoclonal antibody (mAb) for flow cytometry has been reported, this study aimed to develop a novel mAb for mACKR4. Among the established anti-mACKR4 mAbs, A4Mab-1 (rat IgG2b, kappa), A4Mab-2 (rat IgG2b, kappa), and A4Mab-3 (rat IgG2b, kappa) recognized mACKR4-overexpressed Chinese hamster ovary-K1 (CHO/mACKR4) by flow cytometry. The dissociation constant (KD) values of A4Mab-1, A4Mab-2, and A4Mab-3 for CHO/mACKR4 were determined as 6.0 × 10−9 M, 1.3 × 10−8 M, and 1.7 × 10−9 M, respectively. Furthermore, A4Mab-1 and A4Mab-2 could detect mACKR4 by western blotting. These results indicated that A4Mab-1, A4Mab-2, and A4Mab-3 help to detect mACKR4 by flow cytometry and western blotting and obtain the proof of concept in preclinical models.
期刊介绍：
Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.
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