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Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C(44)Mab-108 for Immunohistochemistry
CD44 has been known as a marker of tumor-initiating cells, and plays pro-tumorigenic functions in many cancers. The splicing variants play critical roles in the malignant progression of cancers by promoting stemness, cancer cell invasion or metastasis, and resistance to chemo- and radiotherapy. To u...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10047040/ https://www.ncbi.nlm.nih.gov/pubmed/36975491 http://dx.doi.org/10.3390/cimb45030121 |
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author | Suzuki, Hiroyuki Tanaka, Tomohiro Goto, Nohara Kaneko, Mika K. Kato, Yukinari |
author_facet | Suzuki, Hiroyuki Tanaka, Tomohiro Goto, Nohara Kaneko, Mika K. Kato, Yukinari |
author_sort | Suzuki, Hiroyuki |
collection | PubMed |
description | CD44 has been known as a marker of tumor-initiating cells, and plays pro-tumorigenic functions in many cancers. The splicing variants play critical roles in the malignant progression of cancers by promoting stemness, cancer cell invasion or metastasis, and resistance to chemo- and radiotherapy. To understand each CD44 variant (CD44v) function is essential to know the property of cancers and the establishment of the therapy. However, the function of the variant 4-encoded region has not been elucidated. Therefore, specific monoclonal antibodies (mAbs) against variant 4 are indispensable for basic research, tumor diagnosis, and therapy. In this study, we established anti-CD44 variant 4 (CD44v4) mAbs by immunizing mice with a peptide containing the variant 4-encoded region. We next performed flow cytometry, western blotting, and immunohistochemistry to characterize them. One of the established clones (C(44)Mab-108; IgG(1), kappa) reacted with CD44v3-10-overexpressed Chinese hamster ovary-K1 cells (CHO/CD44v3-10). The K(D) of C(44)Mab-108 for CHO/CD44 v3-10 was 3.4 × 10(−7) M. In western blot analysis, C(44)Mab-108 detected CD44v3-10 in the lysate of CHO/CD44v3-10 cells. Furthermore, C(44)Mab-108 stained formalin-fixed paraffin-embedded (FFPE) oral squamous carcinoma tissues in immunohistochemistry. These results indicated that C(44)Mab-108 is useful to detect CD44v4 in immunohistochemistry using FFPE tissues. |
format | Online Article Text |
id | pubmed-10047040 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-100470402023-03-29 Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C(44)Mab-108 for Immunohistochemistry Suzuki, Hiroyuki Tanaka, Tomohiro Goto, Nohara Kaneko, Mika K. Kato, Yukinari Curr Issues Mol Biol Article CD44 has been known as a marker of tumor-initiating cells, and plays pro-tumorigenic functions in many cancers. The splicing variants play critical roles in the malignant progression of cancers by promoting stemness, cancer cell invasion or metastasis, and resistance to chemo- and radiotherapy. To understand each CD44 variant (CD44v) function is essential to know the property of cancers and the establishment of the therapy. However, the function of the variant 4-encoded region has not been elucidated. Therefore, specific monoclonal antibodies (mAbs) against variant 4 are indispensable for basic research, tumor diagnosis, and therapy. In this study, we established anti-CD44 variant 4 (CD44v4) mAbs by immunizing mice with a peptide containing the variant 4-encoded region. We next performed flow cytometry, western blotting, and immunohistochemistry to characterize them. One of the established clones (C(44)Mab-108; IgG(1), kappa) reacted with CD44v3-10-overexpressed Chinese hamster ovary-K1 cells (CHO/CD44v3-10). The K(D) of C(44)Mab-108 for CHO/CD44 v3-10 was 3.4 × 10(−7) M. In western blot analysis, C(44)Mab-108 detected CD44v3-10 in the lysate of CHO/CD44v3-10 cells. Furthermore, C(44)Mab-108 stained formalin-fixed paraffin-embedded (FFPE) oral squamous carcinoma tissues in immunohistochemistry. These results indicated that C(44)Mab-108 is useful to detect CD44v4 in immunohistochemistry using FFPE tissues. MDPI 2023-02-25 /pmc/articles/PMC10047040/ /pubmed/36975491 http://dx.doi.org/10.3390/cimb45030121 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 Suzuki, Hiroyuki Tanaka, Tomohiro Goto, Nohara Kaneko, Mika K. Kato, Yukinari Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C(44)Mab-108 for Immunohistochemistry |
title | Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C(44)Mab-108 for Immunohistochemistry |
title_full | Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C(44)Mab-108 for Immunohistochemistry |
title_fullStr | Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C(44)Mab-108 for Immunohistochemistry |
title_full_unstemmed | Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C(44)Mab-108 for Immunohistochemistry |
title_short | Development of a Novel Anti-CD44 Variant 4 Monoclonal Antibody C(44)Mab-108 for Immunohistochemistry |
title_sort | development of a novel anti-cd44 variant 4 monoclonal antibody c(44)mab-108 for immunohistochemistry |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10047040/ https://www.ncbi.nlm.nih.gov/pubmed/36975491 http://dx.doi.org/10.3390/cimb45030121 |
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