PNAS-2蛋白单克隆抗体的制备和初步鉴定  

Preparation and Identification of Monoclonal Antibody against PNAS-2 Protein

作　　者：刘佳[1] 陈芳源[1] 王海嵘[1] 钟济华[1] 钟华[1] 韩洁英[1] 欧阳仁荣[1] 

机构地区：[1]上海交通大学医学院附属仁济医院血液科,上海200001

出　　处：《中国实验血液学杂志》2009年第5期1269-1272,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金(编号30670881);上海仁济医院-交通大学基础医学院合作研究基金项目(编号ZD0704)

摘　　要：本研究制备凋亡相关蛋白PNAS-2的单克隆抗体,并对其进行初步鉴定,为PNAS-2蛋白功能研究提供必需工具。以PNAS-2的合成肽为免疫原,通过动物免疫、细胞融合、克隆化制备抗PNAS-2蛋白的单克隆抗体,并用Western blot、免疫荧光、ELISA实验对单克隆抗体的特异性、效价、亚型进行了检测。结果表明:获得了稳定的分泌型杂交瘤细胞株S-31-7,属IgG1λ亚型,腹水效价为1:8000;Western blot测得分子量为28kD的单条特异性条带;免疫荧光实验显示胞浆中有阳性反应。结论:制备的单克隆抗体特异性好、效价高,能够作为深入研究PNAS-2蛋白功能的有利工具。This study was purposed to prepare and primarily identify the specific monoclonal antibodies （McAbs） against the apoptosis related protein PNAS-2 so as to provide the essential tool for study of PNAS-2 function. The McAbs against PNAS-2 were prepared via the immunization of mice, cell fusion and cloning using synthetic peptide of PNAS-2 as immunogen ; the specificity, titer and subtype of McAb were detected by Western blot, ELISA and immunofluorescence. The results showed that the stable hybridoma cell line S-31-7 producing McAbs against PNAS-2 protein was successfully obtained. The immunoglobulin of the McAb was identified to be IGgl h. The titer of ascetic fluid fled McAb were 1 ： 8 000. A single specific band with 28 kD was shown in Western blot test, and the antigen recognized was present in cell cytoplasm by immunofluorescence. In conclusion, the obtained McAb against PNAS-2 displays strong specificity and high titer, which may be applied to the advanced research on PNAS-2 protein.

关 键 词：PNAS-2蛋白 单克隆抗体 白血病 

分 类 号：R733.7[医药卫生—肿瘤]