{"title":"Development of a Novel Anti-Mouse CCR6 Monoclonal Antibody (C<sub>6</sub>Mab-13) by N-Terminal Peptide Immunization.","authors":"Teizo Asano, Tomohiro Tanaka, Hiroyuki Suzuki, Guanjie Li, Ren Nanamiya, Nami Tateyama, Yu Isoda, Yuki Okada, Hiyori Kobayashi, Takeo Yoshikawa, Mika K Kaneko, Yukinari Kato","doi":"10.1089/mab.2022.0021","DOIUrl":null,"url":null,"abstract":"<p><p>The CC chemokine receptor 6 (CCR6) is a G protein-coupled receptor family member that is highly expressed in B lymphocytes, certain subsets of effector and memory T cells, and immature dendritic cells. CCR6 has only one chemokine ligand, CCL20. The CCL20-CCR6 axis has been recognized as a therapeutic target for autoimmune diseases and tumor. This study developed specific monoclonal antibodies (mAbs) against mouse CCR6 (mCCR6) using the peptide immunization method. The established anti-mCCR6 mAb, C<sub>6</sub>Mab-13 (rat IgG<sub>1</sub>, kappa), reacted with mCCR6-overexpressed Chinese hamster ovary-K1 (CHO/mCCR6), and mCCR6-endogenously expressed P388 (mouse lymphoid neoplasma) and J774-1 (mouse macrophage-like) cells in flow cytometry. The dissociation constant (<i>K</i><sub>D</sub>) of C<sub>6</sub>Mab-13 for CHO/mCCR6 cells was determined to be 2.8 × 10<sup>-9</sup> M, indicating that C<sub>6</sub>Mab-13 binds to mCCR6 with high affinity. In summary, C<sub>6</sub>Mab-13 is useful for detecting mCCR6-expressing cells through flow cytometry.</p>","PeriodicalId":53514,"journal":{"name":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","volume":"41 6","pages":"343-349"},"PeriodicalIF":0.0000,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"3","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Monoclonal Antibodies in Immunodiagnosis and Immunotherapy","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1089/mab.2022.0021","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 3
Abstract
The CC chemokine receptor 6 (CCR6) is a G protein-coupled receptor family member that is highly expressed in B lymphocytes, certain subsets of effector and memory T cells, and immature dendritic cells. CCR6 has only one chemokine ligand, CCL20. The CCL20-CCR6 axis has been recognized as a therapeutic target for autoimmune diseases and tumor. This study developed specific monoclonal antibodies (mAbs) against mouse CCR6 (mCCR6) using the peptide immunization method. The established anti-mCCR6 mAb, C6Mab-13 (rat IgG1, kappa), reacted with mCCR6-overexpressed Chinese hamster ovary-K1 (CHO/mCCR6), and mCCR6-endogenously expressed P388 (mouse lymphoid neoplasma) and J774-1 (mouse macrophage-like) cells in flow cytometry. The dissociation constant (KD) of C6Mab-13 for CHO/mCCR6 cells was determined to be 2.8 × 10-9 M, indicating that C6Mab-13 binds to mCCR6 with high affinity. In summary, C6Mab-13 is useful for detecting mCCR6-expressing cells through flow cytometry.