Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Development of a Novel Anti-CD44 Monoclonal Antibody C44Mab-46 for Multiple Applications against Esophageal Squamous Cell Carcinomas

Version 1 : Received: 28 February 2022 / Approved: 1 March 2022 / Online: 1 March 2022 (10:32:33 CET)

How to cite: Goto, N.; Suzuki, H.; Tanaka, T.; Asano, T.; Kaneko, M.K.; Kato, Y. Development of a Novel Anti-CD44 Monoclonal Antibody C44Mab-46 for Multiple Applications against Esophageal Squamous Cell Carcinomas. Preprints 2022, 2022030015 (doi: 10.20944/preprints202203.0015.v1). Goto, N.; Suzuki, H.; Tanaka, T.; Asano, T.; Kaneko, M.K.; Kato, Y. Development of a Novel Anti-CD44 Monoclonal Antibody C44Mab-46 for Multiple Applications against Esophageal Squamous Cell Carcinomas. Preprints 2022, 2022030015 (doi: 10.20944/preprints202203.0015.v1).

Abstract

CD44 is a cell surface glycoprotein, which is widely expressed on normal and cancer cells. CD44 is involved in cell adhesion, migration, proliferation, survival, stemness, and chemo-resistance. Therefore, CD44 is thought to be a promising target for cancer diagnosis and therapy. In this study, we established anti-CD44 monoclonal antibodies (mAbs) by immunizing mice with CD44v3-10 ectodomain and screening using enzyme-linked immunosorbent assay. We then characterized them using flow cytometry, western blotting, and immunohistochemistry. One of the established clones (C44Mab-46; IgG1, kappa) reacted with CD44s-overexpressed Chinese hamster ovary-K1 cells (CHO/CD44s) or esophageal squamous cell carcinoma (ESCC) cell lines (KYSE70 and KYSE770). The KD of C44Mab-46 for CHO/CD44s, KYSE70, and KYSE770 was 1.1×10-8 M, 4.9×10-8 M, and 4.1×10-8 M, respectively. C44Mab-46 detected CD44s of CHO/CD44s and KYSE70, and CD44v of KYSE770 in western blot analysis. Furthermore, C44Mab-46 strongly stained esophageal squamous carcinoma cells in immunohistochemistry using formalin-fixed paraffin-embedded ESCC tissues. Taken together, C44Mab-46 is very useful for detecting CD44 in various applications.

Keywords

CD44; monoclonal antibody; esophageal cancer

Subject

MEDICINE & PHARMACOLOGY, Oncology & Oncogenics

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