title: Monoclonal antibodies for diagnosis of enterovirus 71.
abstract: Enterovirus 71 (EV71), one of the major causative agents of hand, foot, and mouth disease (HFMD), is now recognized as an emerging neurotropic virus in Asia and may cause severe neurologic complications and mortalities. Laboratory diagnosis of EV71 infection must be efficient and accurate, which could be accomplished by various immunoassays. In this study we use a live EV71 isolate, Tainan/4643/98, with genotype C2 as an immunogen to sensitize BALB/c (H-2(d)) mice and then generate the EV71-specific murine monoclonal antibodies. Five hybridoma clones were established and their monoclonal antibodies were characterized. All five clones are applicable in immunofluorescence staining but with different sensitivities-that is, MAbs 22, 24, and 27 were sensitive in IFA detection, and MAbs 22 and 24 were also confirmed in flow cytometry. None of these cross-reacted with coxsackievirus A16 (CVA16) or Echovirus type 6 (ECHO6), but each varied in binding to different EV71 subgenogroups (B1, B4, B5, C2, and C4). Western blot analysis revealed that all of these MAbs reacted with EV71 VP1 capsid proteins, and in addition MAbs 22 and 24 exhibited potent neutralizing activities against EV71 and protected cells from infection. Further, mapping the epitopes for each MAb revealed that only MAb 27 showed positive for the linear epitope DVIESSIGDSVSRAL, which was located at the N-terminus (a.a. 6-20) of EV71 VP1 and highly conserved among all EV71 subgenotypes. Thus, these MAbs may provide valuable tools for the laboratory diagnosis of EV71 infection and for vaccine development.
authors: 1 Key Laboratory of Pain Basic Research and Clinical Therapy, Department of Neurobiology, Medical College of Soochow University , Suzhou, China .
authors: Li
authors: L
authors: Xu
authors: Kao-Jean
authors: KJ
authors: Huang
authors: Tzong-Shiann
authors: TS
authors: Ho
authors: Chia-Chyi
authors: CC
authors: Liu
authors: Ying-Ray
authors: YR
authors: Lee
authors: Ching-Yen
authors: CY
authors: Lin
authors: David
authors: D
authors: Shiuan
authors: Xing-Hong
authors: XH
authors: Jiang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000096:biosynthesis / Q000737:chemistry* / Q000494:pharmacology; D057134:Antibodies, Neutralizing / Q000096:biosynthesis / Q000737:chemistry* / Q000494:pharmacology; D000918:Antibody Specificity; D000998:Antiviral Agents / Q000737:chemistry / Q000494:pharmacology; D015153:Blotting, Western; D045744:Cell Line, Tumor; D017124:Conserved Sequence; D029821:Enterovirus A, Human / Q000187:drug effects / Q000235:genetics / Q000276:immunology*; D004769:Enterovirus Infections / Q000175:diagnosis* / Q000821:virology; D004797:Enzyme-Linked Immunosorbent Assay; D005838:Genotype; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D014764:Viral Proteins / Q000737:chemistry / Q000276:immunology
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D057134:Antibodies, Neutralizing; D000998:Antiviral Agents; D014764:Viral Proteins