Characterization and application of monoclonal antibodies against N protein of SARS-coronavirus.
Severe acute respiratory syndrome-coronavirus (SARS-CoV) causes an infectious disease through respiratory route. Diagnosing the disease effectively and accurately at early stage is essential for preventing the disease transmission and performing antiviral treatment. In this study, we raised monoclonal antibodies (mAbs) against the nucleocapsid (N) protein of SARS-CoV and mapped epitopes by using different truncated N protein fragments. The mapping of those epitopes was valuable for constructing pair-Abs used in serological diagnosis. The results showed that all of the six raised mAbs were divided into two groups recognizing the region of amino acids 249-317 (A group) or 317-395 (B group). This region spanning amino acids 249-395 contains predominant B cell epitopes located at the C-terminus of N protein. One pair-Abs, consisting of N protein-specific rabbit polyclonal antibody and SARS-CoV N protein-specific mAb, was selected to construct a sandwich ELISA-kit. The kit was able to specifically detect SARS-CoV N proteins in serum samples.
Shang
Bo
B
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institute of Biological Sciences, Chinese Academy of Sciences, 320 Yue-Yang Road, Shanghai 200031, China.
Wang
Xiao-Yi
XY
Yuan
Jian-Wei
JW
Vabret
Astrid
A
Wu
Xiao-Dong
XD
Yang
Rui-Fu
RF
Tian
Lin
L
Ji
Yong-Yong
YY
Deubel
Vincent
V
Sun
Bing
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000086462:Coronavirus Nucleocapsid Proteins; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D019590:Nucleocapsid Proteins / Q000276:immunology*; D011817:Rabbits; D012680:Sensitivity and Specificity; D045169:Severe Acute Respiratory Syndrome / Q000097:blood / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000086462:Coronavirus Nucleocapsid Proteins; D000939:Epitopes; D019590:Nucleocapsid Proteins