流式细胞术高敏感抗小鼠HER2单克隆抗体的研制

Tsunenori Ouchida, Hiroyuki Suzuki, Tomohiro Tanaka, M. Kaneko, Y. Kato
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引用次数: 0

摘要

人表皮生长因子受体2 (HER2)在乳腺癌中的过表达是曲妥珠单抗等单克隆抗体(mAb)治疗的重要靶点。由于抗体-药物偶联物曲妥珠单抗-德鲁德康的发展,her2阳性乳腺癌患者的可靶向性已经扩大。为了评估使用抗her2单克隆抗体的发展模式,需要可靠的临床前小鼠模型。因此,应该建立针对小鼠HER2的敏感单克隆抗体(mHER2)。本研究利用基于细胞的免疫和筛选(CBIS)方法开发了抗mher2单克隆抗体。建立的抗mHER2单克隆抗体H2Mab-300(大鼠IgG2b, kappa)和H2Mab-304(大鼠IgG1, kappa)通过流式细胞术与mHER2过表达的中国仓鼠卵巢k1 (CHO/mHER2)和内源性mHER2表达细胞系NMuMG(小鼠乳腺上皮细胞)反应。此外,这些单克隆抗体无法识别mher2敲除的NMuMG细胞。流式细胞术动力学分析表明,H2Mab-300和H2Mab-304对CHO/mHER2的解离常数(KD)分别为1.2 × 10−9 M和1.7 × 10−9 M。H2Mab-300和H2Mab-304对NMuMG的KD值分别为4.9 × 10−10 M和9.0 × 10−10 M。这些结果表明H2Mab-300和H2Mab-304可以应用于流式细胞术检测mHER2,可能有助于在临床前研究中获得概念证明。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Development of Highly Sensitive Anti-Mouse HER2 Monoclonal Antibodies for Flow Cytometry
Overexpression of human epidermal growth factor receptor 2 (HER2) in breast cancer is an important target of monoclonal antibody (mAb) therapy such as trastuzumab. Due to the development of trastuzumab–deruxtecan, an antibody-drug conjugate, the targetable HER2-positive breast cancer patients have been expanded. To evaluate the developing modalities using anti-HER2 mAbs, reliable preclinical mouse models are required. Therefore, sensitive mAbs against mouse HER2 (mHER2) should be established. This study developed anti-mHER2 mAbs using the Cell-Based Immunization and Screening (CBIS) method. The established anti-mHER2 mAbs, H2Mab-300 (rat IgG2b, kappa) and H2Mab-304 (rat IgG1, kappa), reacted with mHER2-overexpressed Chinese hamster ovary-K1 (CHO/mHER2) and endogenously mHER2-expressed cell line, NMuMG (a mouse mammary gland epithelial cell) via flow cytometry. Furthermore, these mAbs never recognized mHER2-knockout NMuMG cells. The kinetic analysis using flow cytometry indicated that the dissociation constant (KD) values of H2Mab-300 and H2Mab-304 for CHO/mHER2 were 1.2 × 10−9 M and 1.7 × 10−9 M, respectively. The KD values of H2Mab-300 and H2Mab-304 for NMuMG were 4.9 × 10−10 M and 9.0 × 10−10 M, respectively. These results indicated that H2Mab-300 and H2Mab-304 could apply to the detection of mHER2 using flow cytometry and may be useful to obtain the proof of concept in preclinical studies.
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来源期刊
Journal of International Translational Medicine
Journal of International Translational Medicine MEDICINE, RESEARCH & EXPERIMENTAL-
自引率
0.00%
发文量
317
审稿时长
8 weeks
期刊介绍: Journal of International Translational Medicine (JITM, ISSN 2227-6394), founded in 2012, is an English academic journal published by Journal of International Translational Medicine Co., Ltd and sponsored by International Fderation of Translational Medicine. JITM is an open access journal freely serving to submit, review, publish, read and download full text and quote. JITM is a quarterly publication with the first issue published in March, 2013, and all articles published in English are compiled and edited by professional graphic designers according to the international compiling and editing standard. All members of the JITM Editorial Board are the famous international specialists in the field of translational medicine who come from twenty different countries and areas such as USA, Britain, France, Germany and so on.
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