Preparation of adeno-associated virus serotype 8 using baculovirus system / 第二军医大学学报

ABSTRACT

Objective: To set up a baculovirus system for preparing recombinant adeno-associated virus serotype 8 (rAAV8) and to preliminarily examine the infection viability of the prepared virus in vitro. Methods: The rAAV8-EGFP was prepared using baculovirus system and was subsequently extracted and purified by high performance liquid chromatography. The viral titer was determined by real-time quantitative PCR and its infection viability for HEK-293 cells was examined by observing the EGFP reporter. Results: Real-time quantitative PCR showed that high titer of rAAV8-EGFP was prepared (1.5×10 13vg/bottle[100 ml medium]) and strong expression of EGFP was observed in HEK-293 cells infected with rAAV8-EGFP. Conclusion: The baculovirus system is capable of producing rAAV8 with high titer and infection viability, paving a way for future research.