Purification and characterization of Epstein-Barr virus gp340/220 produced by a bovine papillomavirus virus expression vector system.
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.
Madej
M
M
Department of Molecular Biology, Paterson Institute for Cancer Research, Manchester, UK.
Conway
M J
MJ
Morgan
A J
AJ
Sweet
J
J
Wallace
L
L
Qualtiere
L F
LF
Arrand
J R
JR
Mackett
M
M
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
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