A lef5-deficient baculovirus expression system with no virion contamination and promoting secretion.

The baculovirus expression system (BEVS) is widely used in biomanufacturing. However, massive late and very late genes are expressed during baculovirus infection of the host cell, and the protein aggregation has a significant negative impact on the intercellular environment of cells and organelle function. Baculovirus particle contamination also hinders the purification of particulate products. In this study, a novel baculovirus vector of deficient-lef5 (Ac-Δlef5) was established to prevent the production of baculovirus particles. It could transduce cells and replicate viral DNA but the expression of the late and very late genes was reduced so that no virus was packed and budded. The Ac-Δlef5 expression system was successfully used to express a secreted protein, thyroid peroxidase (TPO). Compared to wild-type baculovirus (Ac-wt), Ac-Δlef5 delayed the decline in cell viability and prolonged the harvest period from 4 to 6 days after infection. When expressed in the Ac-Δlef5 system, 98% of TPO was secreted extracellularly, which was about 1.8 times that of the Ac-wt system. Meanwhile, the transcription levels of protein folding-related genes were significantly increased. The results show that the Ac-Δlef5 system is a potential novel viral-free baculovirus expression system, which omits the virion removal operation in biomanufacturing.