title: Purification and characterization of Epstein-Barr virus gp340/220 produced by a bovine papillomavirus virus expression vector system.
abstract: Our initial results with a bovine papilloma virus (BPV) vector expression system indicated that we could produce significant amounts of Epstein-Barr virus (EBV) gp340/220 in the supernatant of a mouse fibroblast cell line. We have now extended these findings to show that the truncated version of gp340/220, where the membrane anchor sequence is deleted, is produced even after extended passage of the cells, at a level of approximately 1 mg/4 x 10(8) cells. A simple purification protocol using Sephacryl S300HR and gelatin agarose gives a product which is greater than 90% pure. This product is recognized by anti-gp340 monoclonal antibodies from five different epitope groups and induces antibody that recognizes the authentic gp340/220 and neutralizes EBV in vitro. The purified gp340/220 can be used in ELISA and stimulates the proliferation of T-cell clones specific for gp340/220. These characteristics, together with the fact that BPV-transformed lines have been utilized for the production of pharmaceuticals for use in humans, suggest that this gp340/220 is suitable as a source of antigen for vaccination to prevent EBV infection and related diseases.
authors.affiliation: Department of Molecular Biology, Paterson Institute for Cancer Research, Manchester, UK.
authors.forename: M
authors.initials: M
authors.lastname: Madej
authors.forename: M J
authors.initials: MJ
authors.lastname: Conway
authors.forename: A J
authors.initials: AJ
authors.lastname: Morgan
authors.forename: J
authors.initials: J
authors.lastname: Sweet
authors.forename: L
authors.initials: L
authors.lastname: Wallace
authors.forename: L F
authors.initials: LF
authors.lastname: Qualtiere
authors.forename: J R
authors.initials: JR
authors.lastname: Arrand
authors.forename: M
authors.initials: M
authors.lastname: Mackett
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D000956:Antigens, Viral / Q000737:chemistry / Q000302:isolation & purification*; D052688:Bovine papillomavirus 1 / Q000737:chemistry / Q000235:genetics*; D002999:Clone Cells / Q000276:immunology; D005347:Fibroblasts / Q000737:chemistry; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D008213:Lymphocyte Activation; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000302:isolation & purification*; D014763:Viral Matrix Proteins
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D008565:Membrane Proteins; D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins