兔抗人β_2糖蛋白Ⅰ多克隆抗体的制备与鉴定  

Preparation and identification of rabbit anti-human β_2-GPI antibody

作　　者：黄宏亮[1] 周红[1] 俞颖[2] 李娜[1] 石文霞[1] 王婷[1] 王海波[1] 

机构地区：[1]江苏大学基础医学与医学技术学院,江苏镇江212013 [2]浙江省中医院检测中心,杭州310006

出　　处：《临床检验杂志》2009年第3期192-194,共3页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金资助项目(30370602;30670907);浙江省自然科学基金资助项目(Y206830);浙江省教育厅基金资助项目(20060746)

摘　　要：目的制备兔抗人β2糖蛋白Ⅰ多克隆抗体,并鉴定抗体的特异性及功能活性。方法以纯化的人血浆β2糖蛋白Ⅰ(β2-GPI)免疫新西兰大白兔,获得抗血清;利用Protein A及β2-GPI亲和层析柱纯化血清中总IgG和抗β2-GPI多克隆抗体;用免疫双向扩散试验(IDD)、ELISA和western-blot等方法鉴定抗体。结果IDD及ELISA鉴定抗体效价分别达到16和1×105;western-blot显示纯化的总IgG和抗β2-GPI抗体与β2-GPI蛋白均有特异反应条带;且抗体与β2-GPI复合物能够刺激单核细胞株THP-1表达组织因子(TF)活性。结论成功获得并纯化兔抗人β2-GPI多克隆抗体,为进一步研究β2-GPI/抗β2-GPI复合物在抗磷脂抗体综合征血栓形成机制中的作用建立了基础。Objective To prepare rabbit polyclonal antibody against human β2-GPI and to characterize the specificity and biological functions of the antibocly. Methods A rabbit was immunized with purified-human β2-GPI- The serum whole IgG and anti-β2-GPI antibody were purified by protein A and β2-GPI affinity chromatography respectively. The specificity of the antibody was determined by IDD, ELISA, and Western blotting （ WB）, and the function of the antibody was analyzed. Results The reactive titers of the anti-serum and the whole IgG reached to 1：16 and 1：100 000 with IDD and ELISA respectively. Both the whole IgG and anti-β2-GPI antibody reacted with β2-GPI in WB. The complex of the antibody and β2-GPI induced the monocytic cell line THP-1 to express tissue factor. Conclusions The polyclonal antibody against human β2-GPI has been prepared in the present study, which provides an efficient reagent for studying mechanisms of the thrombosis associated with antiphospholipid syndrome.

关 键 词：抗β2糖蛋白Ⅰ 多克隆抗体 抗磷脂抗体综合征 

分 类 号：Q813[生物学—生物工程]