title: Generation of murine monoclonal antibodies which cross-neutralize human enterovirus genogroup B isolates.
abstract: A live enterovirus 71 (EV71) isolate designated, EV71/E59, with genotype B4 produced in Vero cells and purified over a sucrose gradient was used as the immunogen to generate EV71-specific murine monoclonal antibodies. Four hybridoma clones derived from the fusion of splenocytes of EV71/E59-preimmunized BALB/c (H-2(d)) mice and the NS-1 myeloma cells that exhibit stable growth were selected for detailed characterization. The proof that the hybridomas produced are indeed true independent clones was based on the obervations that they expressed different complementarity-determining regions (CDRs) in their κ light chain genes. Purified ascitic fluids produced by the individual clones reacted against the viral capsid protein, VP1, in Western blot; and recognized distinct sites of a common epitope localized at the C-terminal half of VP1. Each of the monoclonal antibodies exhibited potent neutralizing activities against the immunizing virus strain, as well as two other isolates namely, N0781-TW-01, and N2838, of subgenogroups B4 and B5, respectively, that were found commonly in recent outbreaks in Taiwan. It was also observed the monoclonal antibodies acted cooperatively in neutralizing the EV71/E59 virus.
authors.affiliation: Vaccine Research and Development Center, National Health Research Institutes, Zhunan Township, Miaoli County 350, Taiwan. h.w.chang@nhri.org.tw
authors.forename: Hsuen-Wen
authors.initials: HW
authors.lastname: Chang
authors.forename: Chia-Chyi
authors.initials: CC
authors.lastname: Liu
authors.forename: Min-Han
authors.initials: MH
authors.lastname: Lin
authors.forename: Hui-Min
authors.initials: HM
authors.lastname: Ho
authors.forename: Ya-Ting
authors.initials: YT
authors.lastname: Yang
authors.forename: Yen-Hung
authors.initials: YH
authors.lastname: Chow
authors.forename: Pele
authors.initials: P
authors.lastname: Chong
authors.forename: Charles
authors.initials: C
authors.lastname: Sia
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D057134:Antibodies, Neutralizing / Q000276:immunology* / Q000302:isolation & purification; D002522:Chlorocebus aethiops; D003429:Cross Reactions; D029822:Enterovirus B, Human / Q000276:immunology; D004769:Enterovirus Infections / Q000821:virology; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D013624:Taiwan; D014709:Vero Cells
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing