title: Recombinant glycoprotein E produced in mammalian cells in large-scale as an antigen for varicella-zoster-virus serology.
abstract: A recombinant glycoprotein E (gE) from varicella-zoster virus (VZV) was generated and produced in Chinese Hamster Ovary (CHO) cells, in the development of a specific antigen for analysis of IgG antibodies to VZV. Several stable gE-secreting clones were established and one clone was adapted to growth in serum-free suspension culture. When the cells were cultured in a perfusion bioreactor, gE was secreted into the medium, from where it could be easily purified. The recombinant gE was then evaluated as a serological antigen in ELISA. When compared to a conventional whole virus antigen, the VZV gE showed similar results in ELISA-based seroprevalence studies of 854 samples derived from blood donors, students, ischemic stroke patients and their controls, including samples with border-line results in previous analyses. Eight samples (0.9%) were discordant, all being IgG-negative by the VZV gE ELISA and positive by the whole virus ELISA. The sensitivity and specificity of the VZV gE ELISA were 99.9% and 100%, respectively, compared to 100% and 88.9% for the VZV whole virus ELISA. The elderly subjects showed similar reactivities to both antigens, while VZV gE gave lower signals in the younger cohorts, suggesting that antibodies to gE may increase with age. It was concluded that the recombinant VZV gE from CHO cells was suitable as a serological antigen for the detection of IgG antibodies specific for VZV.
authors.affiliation: Mammalian Protein Expression Core Facility, Sahlgrenska Academy, University of Gothenburg, Box 440, 405 30 Gothenburg, Sweden.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Thomsson
authors.forename: Linn
authors.initials: L
authors.lastname: Persson
authors.forename: Anna
authors.initials: A
authors.lastname: Grahn
authors.forename: Johanna
authors.initials: J
authors.lastname: Snäll
authors.forename: Maria
authors.initials: M
authors.lastname: Ekblad
authors.forename: Eva
authors.initials: E
authors.lastname: Brunhage
authors.forename: Frida
authors.initials: F
authors.lastname: Svensson
authors.forename: Christina
authors.initials: C
authors.lastname: Jern
authors.forename: Gunnar C
authors.initials: GC
authors.lastname: Hansson
authors.forename: Malin
authors.initials: M
authors.lastname: Bäckström
authors.forename: Tomas
authors.initials: T
authors.lastname: Bergström
mesh_terms: D000328:Adult; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D014645:Herpesvirus 3, Human / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D008297:Male; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C056913:glycoprotein E, varicella-zoster virus