Goto Nohara, Suzuki Hiroyuki, Tanaka Tomohiro, Ishikawa Kenichiro, Ouchida Tsunenori, Kaneko Mika K, Kato Yukinari
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.
Antibodies (Basel). 2023 Jun 21;12(3):42. doi: 10.3390/antib12030042.
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.
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。
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