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开发一种新型抗 CD44 单克隆抗体，用于多种治疗食管鳞癌的应用。

Development of a Novel Anti-CD44 Monoclonal Antibody for Multiple Applications against Esophageal Squamous Cell Carcinomas.

机构信息

Department of Molecular Pharmacology, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai 980-8575, Japan.

Ph.D. Program in Human Biology, School of Integrative and Global Majors, University of Tsukuba, Tsukuba 305-8575, Japan.

出版信息

Int J Mol Sci. 2022 May 16;23(10):5535. doi: 10.3390/ijms23105535.

DOI:10.3390/ijms23105535
PMID:35628345
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9146722/
Abstract

CD44 is a cell surface glycoprotein, which is expressed on normal cells, and overexpressed on cancer cells. CD44 is involved in cell adhesion, migration, proliferation, survival, stemness, and chemo-resistance. Therefore, CD44 is thought to be a promising target for cancer diagnosis and therapy. In this study, we established anti-CD44 monoclonal antibodies (mAbs) by immunizing mice with a CD44 variant (CD44v3-10) ectodomain and screening using enzyme-linked immunosorbent assay. We then characterized them using flow cytometry, Western blotting, and immunohistochemistry. One of the established clones (CMab-46; IgG, kappa) reacted with CD44 standard isoform (CD44s)-overexpressed Chinese hamster ovary-K1 cells (CHO/CD44s) or esophageal squamous cell carcinoma (ESCC) cell lines (KYSE70 and KYSE770). The apparent of CMab-46 for CHO/CD44s, KYSE70, and KYSE770 was 1.1 × 10 M, 4.9 × 10 M, and 4.1 × 10 M, respectively. CMab-46 detected CD44s of CHO/CD44s and KYSE70, and CD44 variants of KYSE770 in Western blot analysis. Furthermore, CMab-46 strongly stained the formalin-fixed paraffin-embedded ESCC tissues in immunohistochemistry. Collectively, CMab-46 is very useful for detecting CD44 in various applications.

摘要

CD44 是一种细胞表面糖蛋白，在正常细胞中表达，在癌细胞中过度表达。CD44 参与细胞黏附、迁移、增殖、存活、干性和化疗耐药性。因此，CD44 被认为是癌症诊断和治疗的有前途的靶标。在这项研究中，我们通过用 CD44 变体（CD44v3-10）胞外域免疫小鼠并通过酶联免疫吸附试验筛选来建立抗 CD44 单克隆抗体（mAbs）。然后，我们使用流式细胞术、Western blot 和免疫组织化学对其进行了表征。建立的克隆之一（CMab-46；IgG，kappa）与 CD44 标准同工型（CD44s）过表达的中国仓鼠卵巢-K1 细胞（CHO/CD44s）或食管鳞状细胞癌（ESCC）细胞系（KYSE70 和 KYSE770）反应。CMab-46 对 CHO/CD44s、KYSE70 和 KYSE770 的表观亲和力分别为 1.1×10-7 M、4.9×10-7 M 和 4.1×10-7 M。CMab-46 在 Western blot 分析中检测到 CHO/CD44s 和 KYSE70 的 CD44s 和 KYSE770 的 CD44 变体。此外，CMab-46 在免疫组织化学中强烈染色福尔马林固定石蜡包埋的 ESCC 组织。总之，CMab-46 非常适用于各种应用中 CD44 的检测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b82/9146722/554df6aca8a1/ijms-23-05535-g005.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b82/9146722/8bd88f967a3d/ijms-23-05535-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0b82/9146722/9a54e16e6650/ijms-23-05535-g003.jpg
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