抗ssDNA单克隆抗体的研制和在检测凋亡细胞中的应用

Application of Anti-ssDNA Monoclonal Antibody for the Detection of Apoptotic Cells

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。

In order to set up a new method for the detection of apoptotic cells with anti ssDNA monoclonal antibody,the hybridoma cell clone secreting anti ssDNA antibody was screened and cloned by using hybridoma cell fusion technique The specificity of the monoclonal antibody was identified by competitive ELISA and DNA dot blott ing,and immunohistochemical and immunofluorescent techniques were to be used for the detection of apoptotic cells With the methods in hand,a hybridoma cell line B17 was obtained that secreted anti ssDNA antibody The apoptotic cells could be detected by this monoclonal antibody,and also could be used to differentiate the non apoptotic cells or necrotic cells It concludes that the anti ssDNA monoclonal antibody can be used to detect the apoptotic cells with higher sensitivity and specificity