摘要
从具有高滴度狂犬病毒抗体的多位疫苗注射者采集外周血淋巴细胞,构建人源抗狂犬病毒Fab基因工程抗体文库。用纯化的狂犬aG和CTN株病毒颗粒富集筛选所得Fab噬菌体抗体文库,利用ELISA和间接免疫荧光法IFA鉴定所得人源单克隆抗体Fab段基因的功能特性,并通过序列测定确定所得抗体的轻链和重链的型别,成功获得11株抗狂犬病毒糖蛋白的人源单克隆Fab抗体。将其中5株人源单克隆Fab抗体的轻链和重链分别克隆入全抗体表达载体pAC-L-Fc后转染昆虫Sf9细胞,利用杆状病毒系统实现全抗体的分泌型表达。5株全抗体在体外与狂犬病毒CVS-11株的中和反应中均显示具有狂犬病毒中和活性。人源中和性抗狂犬病毒基因工程全抗体的获得为我国自行生产抗狂犬病单克隆抗体鸡尾酒奠定了物质基础。
A combinatorial human Fab library to the rabies virus was constructed using antibody genes derived from the blood of vaccinated donors.The library were panned and selected on purified rabies virus particles of aG or CTN strain with phage display.Eleven unique human Fab antibodies specific for the rabies virus glycoprotein were obtained by ELISA、IFA and DNA sequences analysis of these antibodies.Among these Fab antibodies,five human Fab antibodies were converted to full-length human IgG antibodies with recombinant baculovirus system.The five full-length human IgG antibodies were tested in vitro for rabies virus neutralization,resulting in all specificities to neutralize the virus.The obtained human anti-rabies antibodies lay the basis for the production of cocktail of anti-rabies monoclonal antibody with chinese intellectual property.
出处
《病毒学报》
CAS
CSCD
北大核心
2010年第4期271-275,共5页
Chinese Journal of Virology
基金
国家863计划(2009AA02Z109)
