TRIP-1抗体的制作及应用  

The Production and Application of TRIP-1 Antibody

作　　者：张宏玲[1,2] 万骏[1,2] 董一辰[1,2] 曹威[1,2] 张新胜[1,2] 张锦勰[1,2] 黄来强[1,2] 

机构地区：[1]清华大学生命科学学院,北京100084 [2]清华大学深圳研究生院生命与健康学部生物医药研究中心和基因与抗体治疗重点实验室,深圳518055

出　　处：《生物技术通报》2013年第3期139-143,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(30570960;30900749);深圳市重点实验室建设与提升计划

摘　　要：旨在制作TGF-β二型受体结合蛋白TRIP-1的抗体,以便推进对该蛋白在TGF-β信号通路以及癌症发生发展中的功能研究。首先克隆出人的TRIP-1基因,构建其原核重组表达载体pET-His-TRIP-1,将其转化至大肠杆菌BL21(DE3)中,诱导获得TRIP-1蛋白,纯化后将其分别免疫新西兰大耳兔和昆明小鼠(KM),免疫4次后取血清,并对抗血清进行效价和亲和力测定。通过Western blot和免疫荧光试验鉴定抗血清特异性。最后,利用proteinG初步纯化抗血清。结果表明,通过重组表达获得抗原蛋白,纯化后免疫新西兰大耳兔和昆明鼠可得到效价较高的抗TRIP-1的多克隆抗体,在稀释倍数较高的情况下仍可以用于Westernblot和免疫荧光试验,且有非常好的特异性。To produce antibody against the TGF-β type Ⅱ receptor binding protein, TRIP-1, contributing to study TRIP-1 ＇s function in TGF-β signaling pathway and cancer development. Human＇s TRIP-1 gene was cloned into the prokaryotic expression vector of pET-His, and then transformed into E. coli BL21 to induce TRIP-1 protein expression. Purified TRIP-1 protein was used to immunize New Zealand rabbits and Kunming mice （ KM ） . The serum was removed and the titer and affinity was determined after immunization 4 times. The antiserum was highly purified by proteinG. Results indicated that the titer of the anti-serum from murine and rabbit are high enough for Western blot and immunofluorescence assay.

关 键 词：原核表达纯化 TRIP-1蛋白 动物免疫 抗血清 抗体鉴定 

分 类 号：Q51[生物学—生物化学]