晚期氧化蛋白产物抗体的制备和鉴定  

Preparation and identification of monoclonal antibody against advanced oxidation protein products

作　　者：田梅[1] 易维京[2] 吴雄飞[1] 余荣杰[1] 

机构地区：[1]第三军医大学西南医院肾科,重庆400038 [2]第三军医大学医学检验系暨药学院,临床生物化学教研室,重庆400038

出　　处：《第三军医大学学报》2009年第20期1973-1976,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30470807)~~

摘　　要：目的制备和鉴定抗人晚期氧化蛋白产物(AOPPs)单克隆抗体(McAb)。方法用次氯酸(HOCl)和纯化的人血白蛋白(HSA)孵育获得AOPPs-HSA,取凝胶层析纯化后的第1峰免疫BALB/c小鼠,采用杂交瘤技术制备抗AOPPs-HSA的McAb,用间接ELISA、Western blot鉴定其特异性及亚类。结果成功筛选出3株稳定分泌抗AOPPs,1株既抗HSA又抗AOPP的McAb杂交瘤细胞株,其分泌抗体亚类3株为IgG1,1株为IgG2b,间接ELISA和Westernblot证明制备的抗体可以特异地和天然或体外制备的AOPPs结合。结论获得抗人AOPPs-HSA的McAb细胞株4株,为进一步研究AOPPs生物学活性及其相关疾病的诊断和预后奠定基础。Objective To prepare and identify the monoclonal antibodies （McAb） against advanced oxidation protein products （AOPPs）. Methods BALB/c mice were immunized with purified AOPPs-human serum albumin （HAS）, and then McAbs against AOPPs-HSA was produced by hybridoma technique. Indirect ELISA and Western blot analysis were used to identify the immunoglobin specificity and subtype. Results Three novel McAbs only against AOPPs-HSA and one against both AOPPs-HSA and HSA were harvested. The subclass of 3 McAbs were determined as IgG1 and 1 McAb as IgG2b. McAbs reacted strongly and specifically to protein of nature and HOCL-treated AOPPs by Western blot analysis. Conclusion The 4 McAbs against AOPPs are prepared and identified successfully, which provide an important tool for biological properties of AOPPs and for its detection clinical laboratory.

关 键 词：晚期氧化蛋白产物 单克隆抗体 制备和鉴定 

分 类 号：R392.33[医药卫生—免疫学] R392.11[医药卫生—基础医学]