title: Expression and purification of human papillomavirus 18 L1 virus-like particle from saccharomyces cerevisiae.
abstract: 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 YEGalpha-HIR525, and transformed into Saccharomyces cerevisiae. 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.
authors: College of Pharmacy, Chung-Ang University, Seoul 156-756, Korea.
authors: Mi-Kyung
authors: MK
authors: Woo
authors: Jung-Mo
authors: JM
authors: An
authors: Jun-Dong
authors: JD
authors: Kim
authors: Sue-Nie
authors: SN
authors: Park
authors: Hong-Jin
authors: HJ
authors: Kim
mesh_terms: D015153:Blotting, Western; D036022:Capsid Proteins / Q000096:biosynthesis / Q000235:genetics; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D052161:Human papillomavirus 18 / Q000302:isolation & purification*; D006801:Humans; D046529:Microscopy, Electron, Transmission; D053918:Papillomavirus Vaccines / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000821:virology*; D014461:Ultracentrifugation; D014614:Vaccines, Synthetic / Q000302:isolation & purification; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D014779:Virus Replication
chemical_list: D036022:Capsid Proteins; D053918:Papillomavirus Vaccines; D014614:Vaccines, Synthetic; D014764:Viral Proteins; C480565:L1 protein, Human papillomavirus type 18