title: Identification of enteroviruses by using monoclonal antibodies against a putative common epitope.
abstract: A common epitope region of enteroviruses was identified by sequence-independent single-primer amplification (SISPA), followed by immunoscreening of 11 cDNA libraries from two Korean enterovirus isolates (echoviruses 7 and 30) and a coxsackievirus B3 (ATCC-VR 30). The putative common epitope region was localized in the N terminus of VP1 when the displayed recombinant proteins from the phages were chased by the convalescent-phase sera. The genomic region encoding the common epitope region was amplified and then expressed by using the vector pGEX-5X-1. The antigenicity of the expressed recombinant protein was identified by Western blotting with guinea pig antisera for six different serotypes of enteroviruses. After successive immunization of mice with the recombinant common epitope protein, splenocytes were extracted and hybridized with P3X63-Ag8-653 cells. A total of 24 hybridomas that produced monoclonal antibodies (MAbs) against the putative common epitope of enteroviruses were selected. Four of these were immunoglobulin G1 isotypes with a kappa light chain. These MAbs recognized 15 Korean endemic serotypes and prototypes of enteroviruses in an indirect immunofluorescence assay. These results suggest that the expressed protein might be a useful antigen for producing group common antibodies and that the use of the MAbs against the putative common epitope of enteroviruses might be a valuable diagnostic tool for rapidly identifying a broad range of enteroviruses.
authors.affiliation: Department of Life Science, College of Natural Science, Chung-Ang University, Seoul 156-756, Korea.
authors.forename: Soo-Youn
authors.initials: SY
authors.lastname: Shin
authors.forename: Ki-Soon
authors.initials: KS
authors.lastname: Kim
authors.forename: Yoon-Sung
authors.initials: YS
authors.lastname: Lee
authors.forename: Yoon-Seok
authors.initials: YS
authors.lastname: Chung
authors.forename: Kwi-Sung
authors.initials: KS
authors.lastname: Park
authors.forename: Doo-Sung
authors.initials: DS
authors.lastname: Cheon
authors.forename: Byoung-Kuk
authors.initials: BK
authors.lastname: Na
authors.forename: Yoonsung
authors.initials: Y
authors.lastname: Kang
authors.forename: Hyang-Min
authors.initials: HM
authors.lastname: Cheong
authors.forename: Youngjoon
authors.initials: Y
authors.lastname: Moon
authors.forename: Jee-Hye
authors.initials: JH
authors.lastname: Choi
authors.forename: Hang-Eui
authors.initials: HE
authors.lastname: Cho
authors.forename: Na-Young
authors.initials: NY
authors.lastname: Min
authors.forename: Jin-Sook
authors.initials: JS
authors.lastname: Son
authors.forename: Young-Hoon
authors.initials: YH
authors.lastname: Park
authors.forename: Youngmee
authors.initials: Y
authors.lastname: Jee
authors.forename: Jae-Deuk
authors.initials: JD
authors.lastname: Yoon
authors.forename: Chul-Yong
authors.initials: CY
authors.lastname: Song
authors.forename: Kwang-Ho
authors.initials: KH
authors.lastname: Lee
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000918:Antibody Specificity; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004770:Enterovirus / Q000145:classification* / Q000276:immunology; D000939:Epitopes / Q000276:immunology*; D005260:Female; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D017931:DNA Primers; D018076:DNA, Complementary; D000939:Epitopes; D014764:Viral Proteins