摘要
将来源于噬菌体抗体库的人源狂犬病毒糖蛋白特异性单抗G10Fab的基因 ,克隆入杆状病毒人源IgG抗体表达载体 ,通过转染将重组质粒导入昆虫细胞 ,以全抗体的形式表达了一株人源抗狂犬病毒基因工程抗体G10。用亲和层析的方法纯化了表达产物 ,经与一株鼠源糖蛋白特异性单抗竞争证实 ,该单克隆抗体特异性识别狂犬病毒糖蛋白 ,亲和力约为 10 -9M。体外中和实验证明 。
Previously,we have reported that the Fab genes of a neutralizing human recombinant antibody G10 to rabies virus gycoprotein were isolated by using phage display technology The heavy and light chain antibody genes were subcloned into Baculovirus human IgG expression cassette vector and linked with IgG Fc portion The recombinant human intact antibody to rabies virus was generated by transfecting the genes into Sf9 insect cells and expressed in Sf9 cells with recombinant baculovirus The expressed recombinant IgG antibody was affinity purified and the specific binding property to the glycoprotein of rabies virus was demonstrated by antigen captured ELISA and competition ELISA The neutralization function of the G10 antibody was investigated and confirmed by animal test
出处
《病毒学报》
CAS
CSCD
北大核心
2001年第3期193-199,共7页
Chinese Journal of Virology
关键词
狂犬病毒
中和抗体
全抗体表达
昆虫细胞表达
体外中和活性
人源抗体
rabies virus
neutralizing antibody
intact antibody
baculovirus-insect expression system
neutralizing ability in vitro
