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为了制备抗单链DNA单克隆抗体和建立检测细胞凋亡的新方法 ,采用杂交瘤细胞融合技术用于筛选分泌抗单链DNA抗体的单克隆杂交瘤细胞株 ,并用斑点印迹和竞争ELISA法鉴定单抗的特异性 ,进而结合免疫组织化学和免疫荧光方法用于检测凋亡细胞。通过筛选得到单克隆杂交瘤细胞株B17,其分泌的抗体与单链DNA结合而与双链DNA无交叉反应 ,用此单抗建立的凋亡细胞检测方法能够检测出凋亡细胞 ,区分非凋亡细胞和坏死细胞。实验结果表明 ,成功地获得一株分泌特异性抗单链DNA抗体的单克隆杂交瘤细胞株 ,以此建立的凋亡细胞检测方法特异性高、敏感性强
In order to prepare anti-single-stranded DNA monoclonal antibodies and to establish a new method for detecting apoptosis, a hybridoma cell fusion technique was used to screen monoclonal hybridoma cell lines secreting anti-single-stranded DNA antibodies and identified by dot blot and competition ELISA Specificity of monoclonal antibodies, and then combined with immunohistochemistry and immunofluorescence assay for the detection of apoptotic cells. The monoclonal antibody hybridoma cell line B17 was screened and its secreted antibody was bound to single-stranded DNA without cross-reaction with double-stranded DNA. The apoptotic cells detected by this monoclonal antibody detection method can detect apoptotic cells and differentiate non-withered Dead cells and necrotic cells. The experimental results show that a monoclonal hybridoma cell line secreting specific anti-single-stranded DNA antibody is successfully obtained, and the detection method of the apoptotic cells established by the method is highly specific and sensitive