Development of a Novel Anti-Mouse CCR6 Monoclonal Antibody (C₆Mab-13) by N-Terminal Peptide Immunization.

Q3 Medicine Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Pub Date : 2022-12-01 DOI:10.1089/mab.2022.0021

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

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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 (K_D) 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.

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新型抗小鼠CCR6单克隆抗体(C6Mab-13)的n端肽免疫制备

CC趋化因子受体6 (CCR6)是G蛋白偶联受体家族成员，在B淋巴细胞、某些效应T细胞和记忆T细胞亚群以及未成熟树突状细胞中高度表达。CCR6只有一个趋化因子配体CCL20。CCL20-CCR6轴已被认为是自身免疫性疾病和肿瘤的治疗靶点。本研究采用多肽免疫方法制备了针对小鼠CCR6 (mCCR6)的特异性单克隆抗体(mab)。建立的抗mCCR6单抗，C6Mab-13(大鼠IgG1, kappa)，在流式细胞术中与mCCR6过表达的中国仓鼠卵巢k1 (CHO/mCCR6)和mCCR6内源性表达的P388(小鼠淋巴样肿瘤)和J774-1(小鼠巨噬细胞样)细胞反应。测定C6Mab-13对CHO/mCCR6细胞的解离常数(KD)为2.8 × 10-9 M，表明C6Mab-13与mCCR6具有较高的亲和力。综上所述，C6Mab-13可用于流式细胞术检测表达mccr6的细胞。

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来源期刊

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy Medicine-Immunology and Allergy

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4.80

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