Epitope analysis of an anti-mouse CCR1 monoclonal antibody S15040E using flow cytometry

IF 2.2 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry and Biophysics Reports Pub Date : 2025-09-13 DOI:10.1016/j.bbrep.2025.102265

Ayaka Okada , Hiroyuki Suzuki , Takao Arimori , Tomohiro Tanaka , Mika K. Kaneko , Yukinari Kato

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引用次数: 0

Abstract

The C–C motif chemokine receptor 1 (CCR1) is widely expressed in various immune cells and plays crucial roles in the maturation and migration of immune cells. CCR1 has been considered an attractive drug target for treating autoimmune diseases and tumors. An anti-mouse CCR1 (mCCR1) monoclonal antibody (clone S15040E) has been used in various in vivo studies to identify mCCR1-positive cells by flow cytometry. However, the binding epitope has not been determined. This study investigated the binding epitope of S15040E using flow cytometry. The mCCR1 extracellular domain-substituted mutant analysis showed that S15040E recognizes the extracellular loop 2 (ECL2, aa 172–197) of mCCR1. Next, alanine (or glycine) scanning was conducted in the ECL2 region. The results revealed that Trp176, Phe178, and Arg181 are essential amino acids for the recognition by S15040E. These results showed the involvement of the ECL2 of mCCR1 in the recognition by S15040E.

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流式细胞术分析抗小鼠CCR1单克隆抗体S15040E的表位

C-C基序趋化因子受体1 （CCR1）广泛表达于多种免疫细胞中，在免疫细胞的成熟和迁移过程中起着至关重要的作用。CCR1被认为是治疗自身免疫性疾病和肿瘤的一个有吸引力的药物靶点。抗小鼠CCR1 （mCCR1）单克隆抗体（克隆S15040E）已在各种体内研究中使用流式细胞术鉴定mCCR1阳性细胞。然而，结合表位尚未确定。本研究利用流式细胞术研究了S15040E的结合表位。mCCR1胞外结构域取代突变体分析显示，S15040E识别mCCR1的胞外环2 （ECL2, aa 172-197）。接下来，在ECL2区域进行丙氨酸（或甘氨酸）扫描。结果表明，Trp176、Phe178和Arg181是S15040E识别的必需氨基酸。这些结果表明mCCR1的ECL2参与了S15040E的识别。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

191

审稿时长

59 days

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.