Production and Prophylactic Efficacy Study of Human Papillomavirus-like Particle Expressing HPV16 L1 Capsid Protein

다국어 초록 (Multilingual Abstract)

To perform the prophylactic study of a vaccine derived from human papillomavirus (HPV) using Balb/c
mice, we produced virus like particles consisting of HPV capsid protein L1 which has been reported
to induce significant humoral and cellular immunity using various animal model systems. In order to
produce HPV16 VLPs, the cDNA of L1 capsid protein in HPV type 16, obtained by polymerase chain
reaction, was inserted into yeast expression vector, YEGa-HIR525 under the control of GAL10 promoter.
The transformation of YEGa-HPV16 L1 was performed into the yeast Saccharomyces cerevisiae
Y2805 by the lithium acetate method and the yeast clone expressing the highest level of L1 capsid protein
of human papillomavirus type 16 was selected by Western blot analysis using anti-HPV16 L1 antibody.
The purification of HPV16 VLP has been performed by the ultracentrifugation and gel-filtration
methods. To validate the vaccine efficacy of the purified HPV16 VLPs and investigate the properties
of HPV16 VLPs to induce humoral immunity, ELISA assay was performed. A significantly increased
production of anti-HPV16 VLP antibodies was observed in sera from immunized mice. The neutralization
activity of antibodies in the sera from the vaccinated mice was demonstrated by a rapid and
simple assay to detect hemagglutihation inhibition activity.