大鼠脑红蛋白(NGB)可溶性原核表达和单克隆抗体制备及鉴定  

Prokaryotic Soluble Expression Preparation and Characterization of Monoclonal Antibody of Rat Neuroglobin

作　　者：孙健伟[1] 韩洪彦[1] 徐文琳[1] 王春丽[1] 王航雁[2] 徐淑玲[3] 杨建萍[3] 张家洁[3] 于凤琴[3] 张成岗[1] 

机构地区：[1]军事医学科学院放射医学研究所,北京100850 [2]中国人民解放军总医院小儿内科,北京100853 [3]河南省郑州市妇幼保健院,郑州450053

出　　处：《中国生物化学与分子生物学报》2003年第6期751-756,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金重大研究计划 (No .90 2 0 80 17);国家海外青年学者合作研究基金 (No .3 0 12 80 10 );国家自然科学基金面上项目 (No .3 0 10 0 0 49;No .3 990 0 0 41;No.3 990 0 0 74);北京市自然科学基金 (No .70 0 2 0 3 0 )资助~~

摘　　要：脑红蛋白 (NGB)是新发现的脑特异的携氧蛋白 .为了对其功能进行深入的研究 ,应用已构建大鼠NGB基因编码区原核表达载体pGEX 4T 2 NGB转化的大肠杆菌BL2 1 ,获得可溶性表达产物 .用谷胱甘肽S 转移酶 (GST)亲和层析柱纯化后作为抗原 ,免疫Balb c小鼠 .通过传统的细胞融合方法制备了抗大鼠NGB的单克隆抗体 ,并对全部 4株单抗进行了亚类和亚型、效价和特异性鉴定 .Neuroglobin (NGB), a recently identified monomer globin in nerve system, is highly affinitive with oxygen similar to myoglobin. After the coding region of rat NGB gene was sub cloned into the prokaryotic expression vector pGEX 4T 2, the recombinant vector was introduced into E. coli BL21 (DE3) cells for efficient expression. The soluble GST fusion protein was purified through GST column and used to immunize Balb/c mouse. The monoclonal antibody was then produced through hybridoma cells prepared following traditional protocol. Finally, four kinds of cell line highly secreting anti NGB monoclonal antibody were successfully screened. The Ig subclass, subtype, titer and specificity of the monoclonal antibody were also identified. The monoclonal antibody of rat NGB will facilitate further functional study of NGB.

关 键 词：大鼠 脑红蛋白 原核可溶性表达 单克隆抗体 

分 类 号：Q786[生物学—分子生物学]