Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

A Novel Anti-CD44 Variant 3 Monoclonal Antibody C44Mab-6 was Established for Multiple Applications

Hiroyuki Suzuki
* ORCID logo ,
Kaishi Kitamura
,
Nohara Goto
,
Kenichiro Ishikawa
,
Tsunenori Ouchida
,
Tomohiro Tanaka
,
Mika K. Kaneko
,
Yukinari Kato
*
Version 1 : Received: 15 April 2023 / Approved: 17 April 2023 / Online: 17 April 2023 (10:44:35 CEST)

A peer-reviewed article of this Preprint also exists.

Suzuki, H.; Kitamura, K.; Goto, N.; Ishikawa, K.; Ouchida, T.; Tanaka, T.; Kaneko, M.K.; Kato, Y. A Novel Anti-CD44 Variant 3 Monoclonal Antibody C44Mab-6 Was Established for Multiple Applications. Int. J. Mol. Sci. 2023, 24, 8411. Suzuki, H.; Kitamura, K.; Goto, N.; Ishikawa, K.; Ouchida, T.; Tanaka, T.; Kaneko, M.K.; Kato, Y. A Novel Anti-CD44 Variant 3 Monoclonal Antibody C44Mab-6 Was Established for Multiple Applications. Int. J. Mol. Sci. 2023, 24, 8411.

Abstract

Cluster of differentiation 44 (CD44) promotes tumor progression through the recruitment of growth factors, and the acquisition of stemness, invasiveness, and drug resistance. CD44 has multiple isoforms including CD44 standard (CD44s) and CD44 variants (CD44v), which have common and unique functions in tumor development. Therefore, the elucidation of each CD44 isoform’s function in tumor is essential for the establishment of CD44-targeting tumor therapy. We have established various anti-CD44s and anti-CD44v monoclonal antibodies (mAbs) through immunization of CD44v3–10-overexpressed cells. In this study, we established C44Mab-6 (IgG1, kappa), which recognized the CD44 variant 3-encoded region (CD44v3) determined by enzyme-linked immunosorbent assay. C44Mab-6 reacted with CD44v3–10-overexpressed Chinese hamster ovary (CHO)-K1 cells (CHO/CD44v3–10) or some cancer cell lines (COLO205 and HSC-3) by flow cytometry. The apparent KD of C44Mab-6 for CHO/CD44v3–10, COLO205, and HSC-3 was 1.5 × 10−9 M, 6.3 × 10−9 M, and 1.9 × 10−9 M, respectively. C44Mab-6 could detect the CD44v3–10 in western blotting, and stained the formalin-fixed paraffin-embedded tumor sections in immunohistochemistry. These results indicate that C44Mab-6 is useful for detecting CD44v3 in various experiments, and expected for the application of tumor diagnosis and therapy.

Keywords

CD44; CD44 variant 3; monoclonal antibody; flow cytometry; immunohistochemistry

Subject

Medicine and Pharmacology, Oncology and Oncogenics

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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