Characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus

• Published in: The Japanese Journal of Veterinary Research Vol. 55; no. 4; pp. 115 - 127
• Main Authors: Kariwa, H.(Hokkaido Univ., Sapporo (Japan)), Noda, H, Nakauchi, M, Ishizuka, M, Hashiguchi, K, Hashimoto, S, Yoshii, K, Asano, A, Agui, T, Kogaki, H, Kurano, Y, Uchida, Y, Fujii, N, Okada, M, Takashima, I
• Format: Journal Article
• Language: English
• Published: Japan Graduate School of Veterinary Medicine Hokkaido University 01.02.2008
• Subjects: Amino Acid Sequence; Amino Acids - chemistry; Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; Antibodies, Viral - chemistry; Antibody Specificity; ANTICORPS MONOCLONAL; ANTICUERPOS MONOCLONALES; Blotting, Western - veterinary; Cercopithecus aethiops; CLONACION MOLECULAR; CLONAGE MOLECULAIRE; Cloning, Molecular; CORONAVIRUS; Coronavirus 229E, Human; Cross Reactions; DIAGNOSTIC TECHNIQUES; ENFERMEDADES RESPIRATORIAS; Epitope Mapping; Fluorescent Antibody Technique, Indirect - veterinary; GENERO HUMANO; GENRE HUMAIN; Humans; MALADIE RESPIRATOIRE; MANKIND; Mice; MOLECULAR CLONING; MONOCLONAL ANTIBODIES; Nucleocapsid Proteins - chemistry; Nucleocapsid Proteins - genetics; Nucleocapsid Proteins - immunology; Rabbits; RESPIRATORY DISEASES; Reverse Transcriptase Polymerase Chain Reaction - veterinary; SARS Virus - immunology; Severe Acute Respiratory Syndrome - diagnosis; Severe Acute Respiratory Syndrome - veterinary; TECNICAS DE DIAGNOSIS; Vero Cells
• ISSN: 0047-1917

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 Qsup(245)TVTKKsup(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.