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Purification and characterization of Epstein-Barr virus gp340/220 produced by a bovine papillomavirus virus expression vector system.

dc.contributor.authorMadej, Monika
dc.contributor.authorConway, Margaret J
dc.contributor.authorMorgan, A J
dc.contributor.authorSweet, J
dc.contributor.authorWallace, L
dc.contributor.authorQualtiere, L F
dc.contributor.authorArrand, John R
dc.contributor.authorMackett, Mike
dc.date.accessioned2010-08-03T08:18:34Z
dc.date.available2010-08-03T08:18:34Z
dc.date.issued1992
dc.identifier.citationPurification and characterization of Epstein-Barr virus gp340/220 produced by a bovine papillomavirus virus expression vector system. 1992, 10 (11):777-82 Vaccineen
dc.identifier.issn0264-410X
dc.identifier.pmid1332270
dc.identifier.doi10.1016/0264-410X(92)90513-J
dc.identifier.urihttp://hdl.handle.net/10541/108874
dc.description.abstractOur 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.
dc.language.isoenen
dc.subject.meshAnimals
dc.subject.meshAntibodies, Viral
dc.subject.meshAntigens, Viral
dc.subject.meshBovine papillomavirus 1
dc.subject.meshClone Cells
dc.subject.meshFibroblasts
dc.subject.meshGene Expression Regulation
dc.subject.meshGenetic Vectors
dc.subject.meshLymphocyte Activation
dc.subject.meshMembrane Proteins
dc.subject.meshMice
dc.subject.meshMice, Inbred BALB C
dc.subject.meshT-Lymphocytes
dc.subject.meshViral Envelope Proteins
dc.subject.meshViral Matrix Proteins
dc.titlePurification and characterization of Epstein-Barr virus gp340/220 produced by a bovine papillomavirus virus expression vector system.en
dc.typeArticleen
dc.contributor.departmentDepartment of Molecular Biology, Paterson Institute for Cancer Research, Manchester, UK.en
dc.identifier.journalVaccineen
html.description.abstractOur 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.


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