OBJECTIVETo express the L1 protein of human papillomavirus type 16 (HPV16) in insect cell suspension culture system.

METHODSOptimized the conditions of suspension culture, recombinant virus amplification and protein expression. Determined the virus tilter by plague analysis and detected the target protein by SDS-PAGE and Western blot; The formation of VLPs by HPV16 L1 protein was observed with TEM.

RESULTSThe Sf9 cells could grow better in suspension culture with seeding density of 5 x 10-5 cell/mL and the maximum expression quantity was obtained by infection of cells with rBacV/HPV16L1 (MOI =10) and harvesting after 72-84 h. HPV16L1 protein could assemble into VLPs in Sf9 cells observed with TEM.

CONCLUSIONThe conditions of cell culture, virus amplification and protein expression were optimized. HPV16 L1 protein could assemble into VLPs in Sf9 cells, which would provide a foundation for further study of the vaccine and diagnosis kits.

Animals ; Capsid Proteins ; biosynthesis ; Cell Proliferation ; Oncogene Proteins, Viral ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Spodoptera ; Suspensions ; Virion ; ultrastructure