Ishikawa, K.; Suzuki, H.; Kaneko, M.K.; Kato, Y. Establishment of a Novel Anti-CD44 Variant 10 Monoclonal Antibody C44Mab-18 for Immunohistochemical Analysis against Oral Squamous Cell Carcinomas. Curr. Issues Mol. Biol.2023, 45, 5248-5262.
Ishikawa, K.; Suzuki, H.; Kaneko, M.K.; Kato, Y. Establishment of a Novel Anti-CD44 Variant 10 Monoclonal Antibody C44Mab-18 for Immunohistochemical Analysis against Oral Squamous Cell Carcinomas. Curr. Issues Mol. Biol. 2023, 45, 5248-5262.
Ishikawa, K.; Suzuki, H.; Kaneko, M.K.; Kato, Y. Establishment of a Novel Anti-CD44 Variant 10 Monoclonal Antibody C44Mab-18 for Immunohistochemical Analysis against Oral Squamous Cell Carcinomas. Curr. Issues Mol. Biol.2023, 45, 5248-5262.
Ishikawa, K.; Suzuki, H.; Kaneko, M.K.; Kato, Y. Establishment of a Novel Anti-CD44 Variant 10 Monoclonal Antibody C44Mab-18 for Immunohistochemical Analysis against Oral Squamous Cell Carcinomas. Curr. Issues Mol. Biol. 2023, 45, 5248-5262.
Abstract
CD44 is known as a cancer stem cell marker of head and neck squamous cell carcinoma (HNSCC) and plays a critical role in cancer malignant progression. Splicing variant isoforms of CD44 (CD44v) are overexpressed in cancers and considered a promising target for cancer therapy. Several monoclonal antibodies (mAbs) against CD44 have been developed by immunizing mice with CD44v3–10-overexpressed cancer cells. In this study, we characterized a novel clone, C44Mab-18 (IgM, kappa). C44Mab-18 reacted with CHO/CD44v3–10, but not with CHO/CD44s by flow cytometry. Enzyme-linked immunosorbent assay revealed that the epitope of C44Mab-18 is determined to be the border sequence between variant 10 and the constant exon 16-encoded sequence. Flow cytometry showed that C44Mab-18 recognizes HSC-3, an oral squamous cell carcinoma(OSCC) cell line. The apparent dissociation constant (KD) of C44Mab-18 for CHO/CD44v3–10 and HSC-3 was determined to be 1.6 × 10−7 M and 1.7 × 10−7 M, respectively. C44Mab-18 detected CD44v3–10, but not CHO/CD44s in western blotting. Furthermore, C44Mab-18 detected endogenous CD44v10 in immunohistochemistry using OSCC tissues. Taken together, C44Mab-18 is useful for detecting CD44v10 in flow cytometry and immunohistochemistry.
Medicine and Pharmacology, Oncology and Oncogenics
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