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EMab-300 Detects Mouse Epidermal Growth Factor Receptor-Expressing Cancer Cell Lines in Flow Cytometry
Nohara Goto1, Hiroyuki Suzuki1,2, Tomohiro Tanaka1
1Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Miyagi, Japan.
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 IgG1, 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 KD of EMab-300 for CHO/mEGFR and NMuMG was 4.3 × 10-8 M and 1.9 × 10-8 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.
Insights
Researchers developed a sensitive monoclonal antibody (mAb), EMab-300, targeting mouse Epidermal Growth Factor Receptor (mEGFR). This new antibody is crucial for preclinical studies of EGFR-targeting cancer therapies in mouse models.
Area of Science:
- Oncology
- Immunology
- Biotechnology
Background:
- Epidermal Growth Factor Receptor (EGFR) signaling is a key driver in various cancers.
- Monoclonal antibody (mAb) therapies targeting human EGFR are clinically approved for certain cancers.
- Development of mouse models is essential for advancing EGFR-targeted mAb therapies.
Purpose of the Study:
- To establish a sensitive and specific monoclonal antibody (mAb) against mouse EGFR (mEGFR).
- To facilitate preclinical research and development of EGFR-targeted therapies using mouse models.
Main Methods:
- Utilized the Cell-Based Immunization and Screening (CBIS) method to develop the mAb.
- Characterized the anti-mEGFR mAb (EMab-300) using flow cytometry on mEGFR-expressing cell lines.
- Performed kinetic analysis to determine antibody binding affinity (KD).
Main Results:
- Developed EMab-300, a rat IgG1 kappa mAb specific for mEGFR.
- EMab-300 demonstrated reactivity with mEGFR-overexpressed and endogenously expressing mouse cell lines.
- Kinetic analysis showed high affinity binding (KD in the range of 10-8 M).
Conclusions:
- EMab-300 is a sensitive and specific mAb for detecting mEGFR via flow cytometry.
- This antibody is valuable for preclinical proof-of-concept studies in cancer research.
- EMab-300 supports the development of novel EGFR-targeted therapies in mouse models.
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