抗汉滩病毒双链抗体重组杆状病毒在昆虫细胞中的表达  

Expression of recombinant baculovirus carrying chimeric gene of a diabody against hantaan virus in insect cells

作　　者：胡刚[1] 徐志凯[1] 张芳琳[1] 吴兴安[1] 阎岩[1] 白文涛[1] 于澜[1] 王海涛[1] 

机构地区：[1]第四军医大学微生物学教研室,西安710032

出　　处：《免疫学杂志》2005年第2期84-87,共4页Immunological Journal

基　　金：国家自然科学基金 (3 9970 70 2 );西安市基金 (YG2 0 0 12 3 )资助项目

摘　　要：目的　应用Bac to Bac杆状病毒表达系统构建具有抗汉滩病毒(HTNV)G2 糖蛋白(GP)鼠源性mAb 3G1与抗HTNV核蛋白(NP)鼠源性mAb 1A8嵌合基因的重组杆状病毒,在昆虫细胞中表达抗HTNV双链抗体并对表达产物进行初步鉴定。方法　构建含有嵌合基因的杆状病毒表达载体ScFv pFBD ScFv’,转化DH10Bac致敏菌,利用其含有的细菌Tn7转座系统将嵌合基因重组至穿梭质粒Bacmid上,快速筛选出含有抗HTNV双链抗体嵌合基因的重组杆状病毒,在昆虫细胞中进行表达,并利用间接免疫荧光试验检测表达产物。结果　构建了含抗HTNV双链抗体嵌合基因之重组杆状病毒,并在昆虫细胞中表达出特异性抗体,该抗体能被兔抗人Cκ抗体所识别并可与HTNVNP和GP特异性结合。结论　成功地在昆虫细胞中表达出抗HTNV双链抗体,且该抗体具有与HTNVNP抗原和GP抗原结合的活性。Objective To express a diabody against hantaan virus (HTNV) in insect cells using the Bac-to-Bac baculovirus expression system, and to preliminarily identify the expression product. Methods The recombinant baculovirus expression vector ScFv-pFBD-ScFv’ carrying chimeric gene of a diabody against HTNV was constructed. The chimeric gene was inserted into shuttle plasmid in E.coli DH10Bac with the help of Tn7 transposition system. The recombinant baculovirus was expressed in insect cells and the expression was identified by immunof- luorescence. Results The recombinant baculovirus containing the chimeric gene was constructed successfully and the diabody could be expressed in insect cells. This diabody could be recognized by the tag of human c κ chain and could combine HTNV nucleoprotein and glycoprotein G 2. Conclusion The diabody against HTNV is constructed and expressed in insect cells successfully. The diabody has the property of binding to HTNV nucleoprotein and glycoprotein.

关 键 词：单链抗体 双链抗体 汉滩病毒 杆状病毒 昆虫细胞 

分 类 号：R392[医药卫生—免疫学]