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 monoclon...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2005-10, Vol.336 (1), p.110-117
Main Authors: Shang, Bo, Wang, Xiao-Yi, Yuan, Jian-Wei, Vabret, Astrid, Wu, Xiao-Dong, Yang, Rui-Fu, Tian, Lin, Ji, Yong-Yong, Deubel, Vincent, Sun, Bing
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - chemistry
Antibodies, Monoclonal - immunology
B cell epitope mapping
Coronavirus Nucleocapsid Proteins
Cross Reactions
Diagnosis
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Epitopes - chemistry
Epitopes - immunology
Human coronaviruses
Humans
Mice
Mice, Inbred BALB C
Monoclonal antibody
Nucleocapsid protein
Nucleocapsid Proteins - immunology
Rabbits
SARS coronavirus
Sensitivity and Specificity
Severe acute respiratory syndrome
Severe Acute Respiratory Syndrome - blood
Severe Acute Respiratory Syndrome - immunology
Truncated N protein fragments
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Summary: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.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2005.08.032