Purification of virus-like particles of recombinant human papillomavirus type 11 major capsid protein L1 from Saccharomyces cerevisiae.
Recombinant major capsid protein, L1 (M(r) = 55,000), of human papillomavirus type 11 was expressed intracellularly at high levels in a galactose-inducible Saccharomyces cerevisiae expression system by an HPV6/11 hybrid gene. The capsid protein self-assembled into virus-like particles (VLPs) and accounted for 15% of the total soluble protein. A purification process was developed that consisted of two main steps: microfiltration and cation-exchange chromatography. The purified VLPs were 98% homogeneous, and the overall purification yield was 10%. The final product was characterized by several analytical methods and was highly immunogenic in mice.
Cook
J C
JC
Department of Virus and Cell Biology, Merck Research Laboratories, West Point, Pennsylvania 19486, USA.
Joyce
J G
JG
George
H A
HA
Schultz
L D
LD
Hurni
W M
WM
Jansen
K U
KU
Hepler
R W
RW
Ip
C
C
Lowe
R S
RS
Keller
P M
PM
Lehman
E D
ED
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000917:Antibody Formation; D015153:Blotting, Western; D002213:Capsid / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D036022:Capsid Proteins; D002852:Chromatography, Ion Exchange; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D027383:Papillomaviridae / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D019065:Virus Assembly
D000596:Amino Acids; D036022:Capsid Proteins; C094776:L1 protein, Human papillomavirus type 11; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins