利用杆状病毒系统获得携带抗体表达基因的双链AAV的研究

Study on Production of Double-strand AAV Carrying Antibody Expressed Gene Using Baculovirus System

研究使用二杆状病毒-昆虫细胞系统获得携带抗体表达基因的8型双链腺相关病毒(scAAV)。首先利用杆状病毒系统获得ssrAAV8-EGFP和scrAAV8-EGFP,实验表明scrAAV8-EGFP比ssrAAV8-EGFP具有更高的转导效率;然后以同样的方法获得scrAAV8-AT7,检测其滴度,并感染HEK293细胞以检测抗体的表达情况。蛋白印迹法检测出阿瓦斯汀(Avastin)重轻链已成功表达,ELISA检测出Avastin在体外的表达量可达770ng/mL。该研究表明利用杆状病毒系统所获得8型scAAV具有生物学活性并能实现外源基因的体外表达。

This study uses baculovirus-insect cell system to produce 8-type double-strand adeno-associated virus （scAAV） carrying antibody expressed gene and first obtains ssrAAV8-EGFP and scrAAV8-EGFP using baculovirus system.The experiment shows that scrAAV8-EGFP has higher transduction efficiency than ssrAAV8-EGFP.Then,it obtains scrAAV8-AT7 with the same method,detects its titer and infects HEK293 cell so as to test antibody expression.It is detected that Avastin heavy and light chains are successfully expressed with western blotting method and that Avastin＇s expression quantity in vitro can reach 770 ng/mL with ELISA.This study shows that 8-type scAAV obtained using baculovirus system has biological activity and can realize expression of exogenous gene in vitro.