A Novel Anti-CD44 Variant 9 Monoclonal Antibody C44Mab-1 Was Developed for Immunohistochemical Analyses against Colorectal Cancers

Current Issues in Molecular Biology, Год журнала: 2023, Номер 45(4), С. 3658 - 3673

Опубликована: Апрель 20, 2023

Cluster of differentiation 44 (CD44) is a type I transmembrane glycoprotein and has been shown to be cell surface marker cancer stem-like cells in various cancers. In particular, the splicing variants CD44 (CD44v) are overexpressed cancers play critical roles stemness, invasiveness, resistance chemotherapy radiotherapy. Therefore, understanding function each CD44v indispensable for CD44-targeting therapy. CD44v9 contains variant 9-encoded region, its expression predicts poor prognosis patients with plays malignant progression tumors. promising target diagnosis Here, we developed sensitive specific monoclonal antibodies (mAbs) against by immunizing mice CD44v3–10-overexpressed Chinese hamster ovary-K1 (CHO/CD44v3–10) cells. We first determined their epitopes using enzyme-linked immunosorbent assay characterized applications as flow cytometry, western blotting, immunohistochemistry. One established clones, C44Mab-1 (IgG1, kappa), reacted peptide indicating that recognizes CD44v9. could recognize CHO/CD44v3–10 or colorectal lines (COLO201 COLO205) cytometric analysis. The apparent dissociation constant (KD) CHO/CD44v3–10, COLO201, COLO205 was 2.5 × 10−8 M, 3.3 6.5 respectively. Furthermore, able detect CD44v3–10 blotting endogenous immunohistochemistry tissues. These results indicated useful detecting not only cytometry but also

Язык: Английский

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Antitumor activities of anti‑CD44 monoclonal antibodies in mouse xenograft models of esophageal cancer

Oncology Reports, Год журнала: 2024, Номер 52(5)

Опубликована: Авг. 29, 2024

CD44 is a type I transmembrane glycoprotein associated with poor prognosis in various solid tumors. Since plays critical role tumor development by regulating cell adhesion, survival, proliferation and stemness, it has been considered target for therapy. Anti‑CD44 monoclonal antibodies (mAbs) have developed applied to antibody‑drug conjugates chimeric antigen receptor‑T Anti-pan‑CD44 mAbs, C₄₄Mab‑5 C₄₄Mab‑46, which recognize both standard (CD44s) variant isoforms were previously developed. The present study generated mouse IgG_2a version of the anti‑pan‑CD44 mAbs (5‑mG_2a C₄₄Mab‑46‑mG_2a) evaluate antitumor activities against CD44‑positive cells. Both 5‑mG_2a C₄₄Mab‑46‑mG_2a recognized CD44s‑overexpressed CHO‑K1 (CHO/CD44s) cells esophageal line (KYSE770) flow cytometry. Furthermore, could activate effector presence CHO/CD44s exhibited complement-dependent cytotoxicity KYSE770 administration significantly suppressed xenograft compared control IgG_2a. These results indicate that exert cancers be promising therapeutic regimen

Язык: Английский

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

International Journal of Molecular Sciences, Год журнала: 2023, Номер 24(9), С. 8411 - 8411

Опубликована: Май 7, 2023

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

Язык: Английский

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Anti-CD44 Variant 10 Monoclonal Antibody Exerts Antitumor Activity in Mouse Xenograft Models of Oral Squamous Cell Carcinomas

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(17), С. 9190 - 9190

Опубликована: Авг. 24, 2024

CD44 regulates cell adhesion, proliferation, survival, and stemness has been considered a tumor therapy target. possesses the shortest standard (CD44s) variety of variant (CD44v) isoforms. Since expression CD44v is restricted in epithelial cells carcinomas compared to CD44s, promising target for monoclonal antibody (mAb) therapy. We previously developed an anti-CD44v10 mAb, C

Язык: Английский

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Establishment of a Novel Anti-CD44 Variant 10 Monoclonal Antibody C44Mab-18 for Immunohistochemical Analysis against Oral Squamous Cell Carcinomas

Current Issues in Molecular Biology, Год журнала: 2023, Номер 45(7), С. 5248 - 5262

Опубликована: Июнь 21, 2023

Head and neck squamous cell carcinoma (HNSCC) is the most common type of head cancer, has been revealed as second-highest expression CD44 in cancers. investigated a cancer stem marker HNSCC plays critical role tumor malignant progression. Especially, splicing variant isoforms (CD44v) are overexpressed cancers considered promising target for diagnosis therapy. We developed monoclonal antibodies (mAbs) against by immunizing mice with CD44v3–10-overexpressed PANC-1 cells. Among established clones, C44Mab-18 (IgM, kappa) reacted CHO/CD44v3–10, but not CHO/CD44s parental CHO-K1 using flow cytometry. The epitope mapping peptides that cover exon-encoded regions recognized border sequence between 10 constant exon 16-encoded sequence. These results suggest recognizes 10-containing CD44v, CD44s. Furthermore, could recognize human oral (OSCC) line, HSC-3, apparent dissociation (KD) CHO/CD44v3–10 HSC-3 was 1.6 × 10−7 M 1.7 M, respectively. detected CD44v3–10 Western blotting, endogenous CD44v10 immunohistochemistry OSCC tissues. indicate useful detecting cytometry immunohistochemistry.

Язык: Английский

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

Опубликована: Апрель 17, 2023

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

Язык: Английский

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Development of a Novel Anti-CD44 Variant 8 Monoclonal Antibody C44Mab-94 against Gastric Carcinomas

Antibodies, Год журнала: 2023, Номер 12(3), С. 45 - 45

Опубликована: Июль 4, 2023

Gastric cancer (GC) is the third leading cause of cancer-related deaths worldwide. GC with peritoneal metastasis exhibits a poor prognosis due to lack effective therapy. A comprehensive analysis malignant ascites identified genomic alterations and significant amplifications driver genes, including CD44. CD44 its splicing variants are overexpressed in tumors, play crucial roles acquisition invasiveness, stemness, resistance treatments. Therefore, development CD44-targeted monoclonal antibodies (mAbs) important for diagnosis In this study, we immunized mice CD44v3-10-overexpressed PANC-1 cells established several dozens clones that produce anti-CD44v3-10 mAbs. One (C44Mab-94; IgG1, kappa) recognized variant-8-encoded region peptide, indicating C44Mab-94 specific mAb CD44v8. Furthermore, could recognize CHO/CD44v3-10 cells, oral squamous cell carcinoma line (HSC-3), or lines (MKN45 NUGC-4) flow cytometric analyses. detect exogenous CD44v3-10 endogenous CD44v8 western blotting stained formalin-fixed paraffin-embedded gastric cells. These results indicate useful detecting variety experimental methods expected become usefully applied

Язык: Английский

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Editorial of Special Issue “Oral Cancer: From Pathophysiology to Novel Therapeutic Approaches”

Biomedicines, Год журнала: 2023, Номер 11(10), С. 2748 - 2748

Опубликована: Окт. 11, 2023

Oral squamous cell carcinoma (OSCC) is a heterogeneous type of malignancy that develops within the oral cavity comprising lips, tongue, mouth floor, gums, and buccal mucosa, with more than 90% arising from lining epithelium [...].

Язык: Английский

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Epitope Mapping of an Anti-CD44v4 Monoclonal Antibody (C₄₄Mab-108) Using Enzyme-Linked Immunosorbent Assay

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, Год журнала: 2024, Номер 43(3), С. 85 - 89

Опубликована: Март 20, 2024

CD44 is a type I transmembrane glycoprotein and possesses various isoforms which are largely classified into standard (CD44s) variant (CD44v) isoforms. Some variant-encoded regions play critical roles in tumor progression. However, the function of 4 (CD44v4)-encoded region has not been fully understood. Using peptide immunization, we developed an anti-CD44v4 monoclonal antibody, C

Язык: Английский

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C₈Mab-21: A Novel Anti-human CCR8 Monoclonal Antibody for Flow Cytometry

Опубликована: Март 19, 2024

C-C motif chemokine receptor-8 (CCR8) belongs to class A of G protein-coupled receptors (GPCRs). CCR8 interacts with the specific ligand CCL1/I-309 in humans, which is produced by various cells, including tumor-associated macrophages and regulatory T cells (Treg). highly expressed on Treg T-helper 2 (TH2) recruited inflammation site implicated allergy, asthma, cancer progression. The CCR8+Treg has been suggested be an important regulator immunosuppressive tumor microenvironment (TME); therefore, it desired develop sensitive monoclonal antibodies (mAbs) for CCR8. This study developed a mAb human (hCCR8), useful flow cytometry employing Cell-Based Immunization Screening (CBIS) method. established anti-hCCR8 mAb, C8Mab-21, (mouse IgM, kappa) reacted hCCR8-overexpressed Chinese hamster ovary-K1 (CHO/hCCR8) TALL-1 (acute lymphoblastic leukemia), CCRF-HSB2 (human T-lymphoblastic natural killer express endogenous hCCR8 cytometry. Furthermore, C8Mab-21 demonstrated moderate binding affinity CHO/hCCR8 dissociation constant 6.5×10-8 M 2.0×10-8 M, respectively. was CBIS method, could tool analyzing hCCR8-related biological response using

Язык: Английский

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