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KMID : 0043320080310020205
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Archives of Pharmacal Research
2008 Volume.31 No. 2 p.205 ~ p.209
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Expression and Purification of Human Papillomavirus 18 L1 Virus-Like Particle from Saccharomyces cerevisiae
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Woo Mi-Kyung
An Jung-Mo
Kim Jun-Dong
Park Sue-Nie
Kim Hong-Jin
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Abstract
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Cervical cancer caused by human papillomavirus (HPV) might be successfully prevented by HPV vaccination and screening. HPV vaccination and HPV serology assays have been investigated using HPV virus-like particles (VLPs). In this study we produced HPV18 L1 VLPs in Saccharomyces cerevisiae and purified them. The HPV18 L1 gene was cloned into the yeast expression vector YEGa-HIR525, and transformed into Saccharomyces cereviaiae. Expression of HPV18 L1 protein was demonstrated by Western blotting. The HPV18 L1 protein was purified by ultracentrifugation, size-exclusion chromatography and cation-exchange chromatography, and was up to 95% pure. We showed by transmission electron microscopy that the purified protein self-assembled into VLPs. These findings should be useful for establishing vaccine efficacy as well as characterizing vaccine candidates, and may provide an international reference standard for HPV serology assays.
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KEYWORD
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HPV18 L1 VLP, Saccharomyces cerevisiae, Expression, Purification
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