서강대학교

초록

Majority of viral vaccines contain inactivated or dead viruses in order to elicit a protective immune response. Virus like Particles that resemble the structure of viruses and elicit strong humoral and cellular reactions, are deemed harmless due to the lack of a viral genome. The Japanese Encephalitis Virus (JEV) vaccine is currently manufactured using animal cells, which poses major health dangers that must be addressed. To this end the JEV Envelope (E) and Membrane (M) sequences were cloned into plant expression vector pHREAC2 generating pSK488 and pSK489 respectively and expressed transiently in Nicotiana benthamiana using Agrobacterium tumefaciens. The expression was optimized for the different Agrobacterium tumefaciens infiltration conditions. For the vector pSK488 and pSK489, 0.6 OD600 with 6 days post-infiltration (dpi) gave highest expression of JEV EM. Ultracentrifugation and Q column chromatography were used in a two-step purification process to obtain the VLP. The molecular structure, which consists of BiP-1 sp coupled to JEV-EM, enables the effective expression of JEV-VLP in Nicotiana benthamiana, but presents certain purification issues that can be overcome with the application of improved purification methods.