EMab-300在流式细胞术中可检测表达小鼠表皮生长因子受体的癌细胞系。

EMab-300 Detects Mouse Epidermal Growth Factor Receptor-Expressing Cancer Cell Lines in Flow Cytometry.

作者信息

Goto Nohara, Suzuki Hiroyuki, Tanaka Tomohiro, Ishikawa Kenichiro, Ouchida Tsunenori, Kaneko Mika K, Kato Yukinari

机构信息

Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Miyagi, Japan.

Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Miyagi, Japan.

出版信息

Antibodies (Basel). 2023 Jun 21;12(3):42. doi: 10.3390/antib12030042.

DOI:10.3390/antib12030042

PMID:37489364

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10366908/

Abstract

Epidermal Growth Factor Receptor (EGFR) overexpression or its mutation mediates the sustaining proliferative signaling, which is an important hallmark of cancer. Human EGFR-targeting monoclonal antibody (mAb) therapy such as cetuximab has been approved for clinical use in patients with colorectal cancers and head and neck squamous cell carcinomas. A reliable preclinical mouse model is essential to further develop the mAb therapy against EGFR. Therefore, sensitive mAbs against mouse EGFR (mEGFR) should be established. In this study, we developed a specific and sensitive mAb for mEGFR using the Cell-Based Immunization and Screening (CBIS) method. The established anti-mEGFR mAb, EMab-300 (rat IgG, kappa), reacted with mEGFR-overexpressed Chinese hamster ovary-K1 (CHO/mEGFR) and endogenously mEGFR-expressed cell lines, including NMuMG (a mouse mammary gland epithelial cell) and Lewis lung carcinoma cells, using flow cytometry. The kinetic analysis using flow cytometry indicated that the of EMab-300 for CHO/mEGFR and NMuMG was 4.3 × 10 M and 1.9 × 10 M, respectively. These results indicated that EMab-300 applies to the detection of mEGFR using flow cytometry and may be useful to obtain the proof of concept in preclinical studies.

摘要

表皮生长因子受体（EGFR）的过表达或其突变介导持续的增殖信号传导，这是癌症的一个重要标志。人源靶向EGFR的单克隆抗体（mAb）疗法，如西妥昔单抗，已被批准用于治疗结直肠癌和头颈部鳞状细胞癌患者。一个可靠的临床前小鼠模型对于进一步开发抗EGFR的单克隆抗体疗法至关重要。因此，应建立针对小鼠EGFR（mEGFR）的敏感单克隆抗体。在本研究中，我们使用基于细胞的免疫和筛选（CBIS）方法开发了一种针对mEGFR的特异性敏感单克隆抗体。所建立的抗mEGFR单克隆抗体EMab-300（大鼠IgG，κ链），通过流式细胞术与过表达mEGFR的中国仓鼠卵巢-K1（CHO/mEGFR）细胞以及内源性表达mEGFR的细胞系（包括NMuMG（一种小鼠乳腺上皮细胞）和Lewis肺癌细胞）发生反应。使用流式细胞术进行的动力学分析表明，EMab-300对CHO/mEGFR和NMuMG的解离常数分别为4.3×10⁻⁹M和1.9×10⁻⁹M。这些结果表明，EMab-300适用于通过流式细胞术检测mEGFR，并且可能有助于在临床前研究中获得概念验证。