重组人瘦素蛋白纯化鉴定及其单克隆抗体制备  

Purification and identification of rh-leptin and preparation of rh-leptin monoclonal antibody

作　　者：谢小强[1] 曹亮[2] 王弦[1] 徐振山[3] 宋礼华[1,3] 

机构地区：[1]安徽医科大学生物化学与分子生物学教研室,合肥230032 [2]安徽大学生命科学学院,合肥230039 [3]安徽安科生物工程集团股份有限公司,合肥230088

出　　处：《生物学杂志》2009年第3期26-28,共3页Journal of Biology

基　　金：安徽省科技攻关项目编号( 07010102247 )

摘　　要：大肠杆菌(E.coli)重组表达获得的重组人瘦素蛋白(rh-leptin),复性、纯化后进行SDS-PAGE电泳和Western-blot印迹杂交鉴定其免疫学活性,免疫小鼠后制备单克隆抗体,结果表明通过对rh-leptin进行复性和纯化,获得了高纯度的具有免疫学活性的rh-leptin蛋白,并获得一株稳定分泌抗rh-leptin单抗的杂交瘤细胞株。瘦素蛋白的纯化及其单克隆抗体的制备,可供瘦素进一步研究应用。The immunological activity of recombined human leptin （rh-leptin） after the protein renaturation and purification was identified by SDS-PAGE and immune mouse with rh-leptin to preparation monoelonal antibody for preparation monoelonal anti- body of reeombined human leptin and purification of reeombined human rh-leptin. Results showed that highly purified rh-leptin with immunological activity by rh-leptin protein renaturation and purification was obtained, and one strain of hybridoma cell can stably secrete MCAB of rh-leptin. By this method, preparation monoelonal antibody of reeombined human leptin and purification of recombined human leptin may be used in further research.

关 键 词：瘦素 蛋白纯化 单克隆抗体 离子交换层析 

分 类 号：R392.33[医药卫生—免疫学]