Expression and purification of human papillomavirus 18 L1 virus-like particle from saccharomyces cerevisiae.
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.
Woo
Mi-Kyung
MK
College of Pharmacy, Chung-Ang University, Seoul 156-756, Korea.
An
Jung-Mo
JM
Kim
Jun-Dong
JD
Park
Sue-Nie
SN
Kim
Hong-Jin
HJ
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
D036022:Capsid Proteins; D053918:Papillomavirus Vaccines; D014614:Vaccines, Synthetic; D014764:Viral Proteins; C480565:L1 protein, Human papillomavirus type 18