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Volume 21 Issue 3

June 2006

Citation: WANG Jian-wei, WANG Yan-bin, CHANG Zhao-rui, YAN Ke-xia, TAN Wen-jie, QU Jian-guo, XIA Ning-shao, RUAN Li, HUNG Tao. Specificity and Epitope Mapping of Four Monoclonal Antibodies against SARS-CoV Nucleocapsid Protein .VIROLOGICA SINICA, 2006, 21(3) : 207-212.

Specificity and Epitope Mapping of Four Monoclonal Antibodies against SARS-CoV Nucleocapsid Protein

WANG Jian-wei ^{1*
,,} ,
WANG Yan-bin ¹ ,
CHANG Zhao-rui ¹ ,
YAN Ke-xia ¹ ,
TAN Wen-jie ¹ ,
QU Jian-guo ¹ ,
XIA Ning-shao ^2* ,
RUAN Li ¹ ,
HUNG Tao ¹

1.
1. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China
2.
The Research Center for Molecular Virology of Fujian Province, The Key Laboratory of Ministry of Education for Cell Biology and Tumor Cell Engineering, Xiamen University, Xiamen 361005, China

Corresponding author: WANG Jian-wei,
Available online: 20 May 2006

Abstract

In order to characterize the specificity of the monoclonal antibodies (McAbs) against SARS- associated coronavirus (SARS-CoV) nucleocapsid protein and to identify the epitopes recognized by the McAbs，the nucleocapsid proteins of human coronavirus OC43 (HCoV-OC43) and 229E (HCoV-229E) was expressed in E.coli. The specificities of four McAbs (1-1C2, 2-2E5, 1-1D6, and 2-8F11) were examined by Western blotting as well as indirect fluorescence assay. Twelve different recombinant truncated N proteins were then used to identify the epitopes recognized by the McAbs by Western blotting. The results showed that: (1) McAbs 1-1C2, 2-2E5, and 1-1D6 recognized neither human coronviruses HCoV-OC43 and HCoV-229E nor their nucleocapsid proteins, implying a possible specificity of these 3 McAbs to SARS-CoV; (2)The epitopes recognized by McAbs 2-8F11, 2-2E5, and 1-1D6 were located between the 30th and 60th amino acid (a.a.) residues of the SARS-CoV N protein, while the epitopes recognized by the McAb1-1C2 were located between 170th and 184 th a.a. residues. The identification of the specific epitopes of SARS-CoV N protein is paramount for the immunological characterization, the development of accurate immunological assay as well as for exploring the pathogenesis of SARS-CoV.
- SARS-associated coronavirus (SARS-CoV)
- , Nucleocapsid protein
- , Monoclonal antibody
- , Epitope

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Specificity and Epitope Mapping of Four Monoclonal Antibodies against SARS-CoV Nucleocapsid Protein

Corresponding author: WANG Jian-wei,

1. 1. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China
2. The Research Center for Molecular Virology of Fujian Province, The Key Laboratory of Ministry of Education for Cell Biology and Tumor Cell Engineering, Xiamen University, Xiamen 361005, China

Abstract: In order to characterize the specificity of the monoclonal antibodies (McAbs) against SARS- associated coronavirus (SARS-CoV) nucleocapsid protein and to identify the epitopes recognized by the McAbs，the nucleocapsid proteins of human coronavirus OC43 (HCoV-OC43) and 229E (HCoV-229E) was expressed in E.coli. The specificities of four McAbs (1-1C2, 2-2E5, 1-1D6, and 2-8F11) were examined by Western blotting as well as indirect fluorescence assay. Twelve different recombinant truncated N proteins were then used to identify the epitopes recognized by the McAbs by Western blotting. The results showed that: (1) McAbs 1-1C2, 2-2E5, and 1-1D6 recognized neither human coronviruses HCoV-OC43 and HCoV-229E nor their nucleocapsid proteins, implying a possible specificity of these 3 McAbs to SARS-CoV; (2)The epitopes recognized by McAbs 2-8F11, 2-2E5, and 1-1D6 were located between the 30th and 60th amino acid (a.a.) residues of the SARS-CoV N protein, while the epitopes recognized by the McAb1-1C2 were located between 170th and 184 th a.a. residues. The identification of the specific epitopes of SARS-CoV N protein is paramount for the immunological characterization, the development of accurate immunological assay as well as for exploring the pathogenesis of SARS-CoV.

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