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Article in Chinese | WPRIM (Western Pacific) | ID: wpr-248756

ABSTRACT

<p><b>OBJECTIVE</b>To express the L1 protein of human papillomavirus type 16 (HPV16) in insect cell suspension culture system.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>


Subject(s)
Animals , Capsid Proteins , Cell Proliferation , Oncogene Proteins, Viral , Recombinant Proteins , Spodoptera , Suspensions , Virion
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