title: Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus.
abstract: The sudden emergence of severe acute respiratory syndrome (SARS) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. As no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a SARS epidemic is the rapid diagnosis and isolation of SARS patients. To establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (NP) of SARS-coronavirus (SARS-CoV). On immunofluorescent antibody assay and Western blotting analysis, none of the monoclonal antibodies showed cross-reactivity to authentic and recombinant NPs of human coronavirus (HCoV) 229E strain. To determine the region on the NP molecule where the monoclonal antibodies bind, we generated four truncated recombinant NPs and analyzed the reactivity between monoclonal antibodies and truncated NPs. Two monoclonal antibodies reacted with a truncated NP covering from amino acid residues 111 to 230, and seven reacted with another truncated NP covering from amino acid residues 221 to 340. Epitope mapping analysis indicated that monoclonal antibody SN5-25 recognized the amino acid sequence Q(245)TVTKK(250) On SARS-NP. Within the epitope, Q245, T246, V247, K249, and K250 appeared to form an essential motif for monoclonal antibody SN5-25 to bind. The information about binding sites and epitopes of monoclonal antibodies may be useful for the development of new diagnostic methods for SARS and for analyzing the function of N protein of SARS-CoV.
authors: Laboratory of Public Health, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan. kariwa@vetemed.hokudai.ac.jp
authors: Hiroaki
authors: H
authors: Kariwa
authors: Hiroshi
authors: H
authors: Noda
authors: Mina
authors: M
authors: Nakauchi
authors: Mariko
authors: M
authors: Ishizuka
authors: Kazuaki
authors: K
authors: Hashiguchi
authors: Shingo
authors: S
authors: Hashimoto
authors: Kentaro
authors: K
authors: Yoshii
authors: Atsushi
authors: A
authors: Asano
authors: Takashi
authors: T
authors: Agui
authors: Hiroyuki
authors: H
authors: Kogaki
authors: Yoshihiro
authors: Y
authors: Kurano
authors: Yoshiaki
authors: Y
authors: Uchida
authors: Nobuyuki
authors: N
authors: Fujii
authors: Masahisa
authors: M
authors: Okada
authors: Ikuo
authors: I
authors: Takashima
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000914:Antibodies, Viral / Q000737:chemistry; D000918:Antibody Specificity; D015153:Blotting, Western / Q000662:veterinary; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D028941:Coronavirus 229E, Human; D000086462:Coronavirus Nucleocapsid Proteins; D003429:Cross Reactions; D018604:Epitope Mapping; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D006801:Humans; D051379:Mice; D019590:Nucleocapsid Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis / Q000662:veterinary*; D014709:Vero Cells
chemical_list: D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins