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Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C(44)Mab-9 for Multiple Applications against Colorectal Carcinomas

CD44 is a cell surface glycoprotein, and its isoforms are produced by the alternative splicing with the standard and variant exons. The CD44 variant exon-containing isoforms (CD44v) are overexpressed in carcinomas. CD44v6 is one of the CD44v, and its overexpression predicts poor prognosis in colorec...

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Autores principales: Ejima, Ryo, Suzuki, Hiroyuki, Tanaka, Tomohiro, Asano, Teizo, Kaneko, Mika K., Kato, Yukinari
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9965047/
https://www.ncbi.nlm.nih.gov/pubmed/36835416
http://dx.doi.org/10.3390/ijms24044007
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author Ejima, Ryo
Suzuki, Hiroyuki
Tanaka, Tomohiro
Asano, Teizo
Kaneko, Mika K.
Kato, Yukinari
author_facet Ejima, Ryo
Suzuki, Hiroyuki
Tanaka, Tomohiro
Asano, Teizo
Kaneko, Mika K.
Kato, Yukinari
author_sort Ejima, Ryo
collection PubMed
description CD44 is a cell surface glycoprotein, and its isoforms are produced by the alternative splicing with the standard and variant exons. The CD44 variant exon-containing isoforms (CD44v) are overexpressed in carcinomas. CD44v6 is one of the CD44v, and its overexpression predicts poor prognosis in colorectal cancer (CRC) patients. CD44v6 plays critical roles in CRC adhesion, proliferation, stemness, invasiveness, and chemoresistance. Therefore, CD44v6 is a promising target for cancer diagnosis and therapy for CRC. In this study, we established anti-CD44 monoclonal antibodies (mAbs) by immunizing mice with CD44v3-10-overexpressed Chinese hamster ovary (CHO)-K1 cells. We then characterized them using enzyme-linked immunosorbent assay, flow cytometry, western blotting, and immunohistochemistry. One of the established clones (C(44)Mab-9; IgG(1), kappa) reacted with a peptide of the variant 6-encoded region, indicating that C(44)Mab-9 recognizes CD44v6. Furthermore, C(44)Mab-9 reacted with CHO/CD44v3-10 cells or CRC cell lines (COLO201 and COLO205) by flow cytometry. The apparent dissociation constant (K(D)) of C(44)Mab-9 for CHO/CD44v3-10, COLO201, and COLO205 was 8.1 × 10(−9) M, 1.7 × 10(−8) M, and 2.3 × 10(−8) M, respectively. C(44)Mab-9 detected the CD44v3-10 in western blotting, and partially stained the formalin-fixed paraffin-embedded CRC tissues in immunohistochemistry. Collectively, C(44)Mab-9 is useful for detecting CD44v6 in various applications.
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spelling pubmed-99650472023-02-26 Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C(44)Mab-9 for Multiple Applications against Colorectal Carcinomas Ejima, Ryo Suzuki, Hiroyuki Tanaka, Tomohiro Asano, Teizo Kaneko, Mika K. Kato, Yukinari Int J Mol Sci Article CD44 is a cell surface glycoprotein, and its isoforms are produced by the alternative splicing with the standard and variant exons. The CD44 variant exon-containing isoforms (CD44v) are overexpressed in carcinomas. CD44v6 is one of the CD44v, and its overexpression predicts poor prognosis in colorectal cancer (CRC) patients. CD44v6 plays critical roles in CRC adhesion, proliferation, stemness, invasiveness, and chemoresistance. Therefore, CD44v6 is a promising target for cancer diagnosis and therapy for CRC. In this study, we established anti-CD44 monoclonal antibodies (mAbs) by immunizing mice with CD44v3-10-overexpressed Chinese hamster ovary (CHO)-K1 cells. We then characterized them using enzyme-linked immunosorbent assay, flow cytometry, western blotting, and immunohistochemistry. One of the established clones (C(44)Mab-9; IgG(1), kappa) reacted with a peptide of the variant 6-encoded region, indicating that C(44)Mab-9 recognizes CD44v6. Furthermore, C(44)Mab-9 reacted with CHO/CD44v3-10 cells or CRC cell lines (COLO201 and COLO205) by flow cytometry. The apparent dissociation constant (K(D)) of C(44)Mab-9 for CHO/CD44v3-10, COLO201, and COLO205 was 8.1 × 10(−9) M, 1.7 × 10(−8) M, and 2.3 × 10(−8) M, respectively. C(44)Mab-9 detected the CD44v3-10 in western blotting, and partially stained the formalin-fixed paraffin-embedded CRC tissues in immunohistochemistry. Collectively, C(44)Mab-9 is useful for detecting CD44v6 in various applications. MDPI 2023-02-16 /pmc/articles/PMC9965047/ /pubmed/36835416 http://dx.doi.org/10.3390/ijms24044007 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Ejima, Ryo
Suzuki, Hiroyuki
Tanaka, Tomohiro
Asano, Teizo
Kaneko, Mika K.
Kato, Yukinari
Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C(44)Mab-9 for Multiple Applications against Colorectal Carcinomas
title Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C(44)Mab-9 for Multiple Applications against Colorectal Carcinomas
title_full Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C(44)Mab-9 for Multiple Applications against Colorectal Carcinomas
title_fullStr Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C(44)Mab-9 for Multiple Applications against Colorectal Carcinomas
title_full_unstemmed Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C(44)Mab-9 for Multiple Applications against Colorectal Carcinomas
title_short Development of a Novel Anti-CD44 Variant 6 Monoclonal Antibody C(44)Mab-9 for Multiple Applications against Colorectal Carcinomas
title_sort development of a novel anti-cd44 variant 6 monoclonal antibody c(44)mab-9 for multiple applications against colorectal carcinomas
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9965047/
https://www.ncbi.nlm.nih.gov/pubmed/36835416
http://dx.doi.org/10.3390/ijms24044007
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