Development of Highly Sensitive Anti-Mouse HER2 Monoclonal Antibodies for Flow Cytometry DOI Creative Commons
Tsunenori Ouchida, Hiroyuki Suzuki, Tomohiro Tanaka

et al.

International Journal of Translational Medicine, Journal Year: 2023, Volume and Issue: 3(3), P. 310 - 320

Published: Aug. 10, 2023

Overexpression of human epidermal growth factor receptor 2 (HER2) in breast cancer is an important target monoclonal antibody (mAb) therapy such as trastuzumab. Due to the development trastuzumab–deruxtecan, antibody-drug conjugate, targetable HER2-positive patients have been expanded. To evaluate developing modalities using anti-HER2 mAbs, reliable preclinical mouse models are required. Therefore, sensitive mAbs against HER2 (mHER2) should be established. This study developed anti-mHER2 Cell-Based Immunization and Screening (CBIS) method. The established H2Mab-300 (rat IgG2b, kappa) H2Mab-304 IgG1, kappa), reacted with mHER2-overexpressed Chinese hamster ovary-K1 (CHO/mHER2) endogenously mHER2-expressed cell line, NMuMG (a mammary gland epithelial cell) via flow cytometry. Furthermore, these never recognized mHER2-knockout cells. kinetic analysis cytometry indicated that dissociation constant (KD) values for CHO/mHER2 were 1.2 × 10−9 M 1.7 M, respectively. KD 4.9 10−10 9.0 These results could apply detection mHER2 may useful obtain proof concept studies.

Language: Английский

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

Nohara Goto,

Hiroyuki Suzuki, Tomohiro Tanaka

et al.

Published: May 9, 2023

The Epidermal Growth Factor Receptor (EGFR) overexpression or its mutation mediates the sustaining proliferative signaling in cancers. Human EGFR-targeting monoclonal antibody (mAb) therapy such as cetuximab has been approved for clinical use patients with colorectal cancers and head neck squamous cell carcinomas. A reliable preclinical mouse model is essential to further develop mAb against EGFR. However, a EGFR (mEGFR) flow cytometry not established. In this study, we developed specific sensitive mEGFR using Cell-Based Immunization Screening (CBIS) method. established anti-mEGFR mAb, EMab-300 (rat IgG1, kappa) reacted mEGFR-overexpressed Chinese hamster ovary-K1 (CHO/mEGFR) endogenously mEGFR-expressed lines, including NMuMG (a mammary gland epithelial cell) Lewis lung carcinoma cells by cytometry. kinetic analysis indicated that dissociation constant (KD) of CHO/mEGFR was 4.3 × 10−8 M 1.9 M, respectively. These results applies detection cytometry, expected study.

Language: Английский

Citations

2
Development of Highly Sensitive Anti-Mouse HER2 Monoclonal Antibodies for Flow Cytometry DOI Creative Commons
Tsunenori Ouchida, Hiroyuki Suzuki, Tomohiro Tanaka

et al.

International Journal of Translational Medicine, Journal Year: 2023, Volume and Issue: 3(3), P. 310 - 320

Published: Aug. 10, 2023

Overexpression of human epidermal growth factor receptor 2 (HER2) in breast cancer is an important target monoclonal antibody (mAb) therapy such as trastuzumab. Due to the development trastuzumab–deruxtecan, antibody-drug conjugate, targetable HER2-positive patients have been expanded. To evaluate developing modalities using anti-HER2 mAbs, reliable preclinical mouse models are required. Therefore, sensitive mAbs against HER2 (mHER2) should be established. This study developed anti-mHER2 Cell-Based Immunization and Screening (CBIS) method. The established H2Mab-300 (rat IgG2b, kappa) H2Mab-304 IgG1, kappa), reacted with mHER2-overexpressed Chinese hamster ovary-K1 (CHO/mHER2) endogenously mHER2-expressed cell line, NMuMG (a mammary gland epithelial cell) via flow cytometry. Furthermore, these never recognized mHER2-knockout cells. kinetic analysis cytometry indicated that dissociation constant (KD) values for CHO/mHER2 were 1.2 × 10−9 M 1.7 M, respectively. KD 4.9 10−10 9.0 These results could apply detection mHER2 may useful obtain proof concept studies.

Language: Английский

Citations

0