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Published on: March 8, 2012
[Expression of HPV16 L1 protein in insect cell suspension culture system]
Cong Han1, Yan Wang, Quing-long Shang
1Department of Microbiology and Human Parasitology, Harbin Medical University, Harbin 150081, China.
Objective:
To express the L1 protein of human papillomavirus type 16 (HPV16) in insect cell suspension culture system.
Methods:
Optimized 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.
Results:
The 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.
Conclusion:
The 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.
