Ouchida, T.; Suzuki, H.; Tanaka, T.; Kaneko, M.K.; Kato, Y. Development of Highly Sensitive Anti-Mouse HER2 Monoclonal Antibodies for Flow Cytometry. Int. J. Transl. Med.2023, 3, 310-320.
Ouchida, T.; Suzuki, H.; Tanaka, T.; Kaneko, M.K.; Kato, Y. Development of Highly Sensitive Anti-Mouse HER2 Monoclonal Antibodies for Flow Cytometry. Int. J. Transl. Med. 2023, 3, 310-320.
Ouchida, T.; Suzuki, H.; Tanaka, T.; Kaneko, M.K.; Kato, Y. Development of Highly Sensitive Anti-Mouse HER2 Monoclonal Antibodies for Flow Cytometry. Int. J. Transl. Med.2023, 3, 310-320.
Ouchida, T.; Suzuki, H.; Tanaka, T.; Kaneko, M.K.; Kato, Y. Development of Highly Sensitive Anti-Mouse HER2 Monoclonal Antibodies for Flow Cytometry. Int. J. Transl. Med. 2023, 3, 310-320.
Abstract
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 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 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) by flow cytometry. Furthermore, these mAbs never recognized mHER2-knockout NMuMG cells. The kinetic analysis using flow cytometry indicated that the dissociation constant (KD) of H2Mab-300 and H2Mab-304 for CHO/mHER2 was 1.2 × 10−9 M and 1.7 × 10−9 M, respectively. The KD of H2Mab-300 and H2Mab-304 for NMuMG was 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.
Keywords
mouse HER2; monoclonal antibody; cell-based Immunization and screening; CBIS
Subject
Medicine and Pharmacology, Oncology and Oncogenics
Copyright:
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