Establishment of C20Mab-11, a novel anti-CD20 monoclonal antibody, for the detection of B cells
Establishment of C20Mab-11, a novel anti-CD20 monoclonal antibody, for the detection of B cells
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DOI：
10.3892/ol.2020.11753
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发表时间：
2020-08-01
期刊：
影响因子：
2.9
通讯作者：
Kato, Yukinari
中科院分区：
文献类型：
--
作者：
Furusawa, Yoshikazu;Kaneko, Mika Kato;Kato, Yukinari
CD20是几种B淋巴细胞抗原之一，已被证明是检测和治疗B细胞淋巴瘤的有效靶点。用于诊断B细胞淋巴瘤的每一种应用都需要敏感和特异的单抗。虽然已经建立了许多抗CD20的单抗，但可用于抗CD20的应用类型有限。在这项研究中，我们旨在利用基于细胞的免疫和筛选(CBIS)方法，建立新型的抗CD20单抗，用于流式细胞术、蛋白质印迹和免疫组织化学分析等广泛应用。其中一种已建立的单抗C(20)Mab-11(IgM，kappa)在CHO-K1或LN229细胞系中检测到CD20的过度表达，表明C(20)Mab-11是CD20特异性的。在Western印迹分析中，C(20)Mab-11不仅检测到CD20在CHO-K1和LN229细胞中的高表达，而且在Ball-1和Raji细胞中也检测到CD20的高表达，并且具有敏感性和特异性。免疫组织化学分析表明，C(20)Mab-11在淋巴滤泡和B细胞淋巴瘤中呈强阳性。这些结果表明，CBIS法制备的C(20)Mab-11可用于流式细胞术、免疫印迹和免疫组织化学分析检测淋巴瘤组织中CD20的表达，并有望用于B细胞淋巴瘤的治疗。
CD20 is one of several B-lymphocyte antigens that has been shown to be an effective target for the detection and treatment of B-cell lymphomas. Sensitive and specific monoclonal antibodies (mAbs) are required for every application used for the diagnosis of B-cell lymphoma. Although many anti-CD20 mAbs have been established, the types of applications, those anti-CD20 can be used in, are limited. In this study, we aimed to establish novel anti-CD20 mAbs to be used for broad applications, such as flow cytometry, western blot, and immunohistochemical analyses, using the Cell-Based Immunization and Screening (CBIS) method. One of the established mAbs, C(20)Mab-11 (IgM, kappa), detected overexpression of CD20 in CHO-K1 or LN229 cell lines, indicating that C(20)Mab-11 is specific for CD20. In western blot analyses, C(20)Mab-11 detected not only overexpression of CD20 in CHO-K1 or LN229 cell lines, but also CD20 of BALL-1 and Raji cells with both sensitivity and specificity. Furthermore, C(20)Mab-11 strongly stained B cells of the lymph follicle and B cell lymphomas in immunohistochemical analyses. These results indicate that C(20)Mab-11 develped by CBIS method, is useful for the detection of CD20 in lymphoma tissues by flow cytometry, western blot, and immunohistochemical analyses and potentially could be beneficial for the treatment of B cell lymphomas.