Purification of virus-like particles of recombinant human papillomavirus type 11 major capsid protein L1 from Saccharomyces cerevisiae

Protein Expr Purif. 1999 Dec;17(3):477-84. doi: 10.1006/prep.1999.1155.

Abstract

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.

MeSH terms

  • Amino Acids / analysis
  • Animals
  • Antibody Formation
  • Blotting, Western
  • Capsid / biosynthesis*
  • Capsid / chemistry
  • Capsid / immunology
  • Capsid / isolation & purification
  • Capsid Proteins
  • Chromatography, Ion Exchange
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Electron
  • Oncogene Proteins, Viral / biosynthesis*
  • Oncogene Proteins, Viral / chemistry
  • Oncogene Proteins, Viral / immunology
  • Oncogene Proteins, Viral / isolation & purification
  • Papillomaviridae / chemistry*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification
  • Saccharomyces cerevisiae / metabolism*
  • Virus Assembly

Substances

  • Amino Acids
  • Capsid Proteins
  • L1 protein, Human papillomavirus type 11
  • Oncogene Proteins, Viral
  • Recombinant Proteins