The Epitope Study on the SARS-CoV Nucleocapsid Protein

Li, Shuting; Lin, Liang; Wang, Hao; Yin, Jianning; Ren, Yan; Zhao, Zhe; Wen, Jie; Zhou, Cuiqi; Zhang, Xumin; Li, Xiaolei; Wang, Jingqiang; Zhou, Zhengfeng; Liu, Jinxiu; Shao, Jianmin; Lei, Tingting; Fang, Jianqiu; Xu, Ningzhi; Liu, Siqi

The nucleocapsid protein (N protein) has been found to be an antigenic protein in a number of coronaviruses. Whether the N protein in severe acute respiratory syndrome-associated coronavirus (SARS-CoV) is antigenic remains to be elucidated. Using Western blot and Enzyme-linked Immunosorbent Assay (ELISA), the recombinant N proteins and the synthesized peptides derived from the N protein were screened in sera from SARS patients. All patient sera in this study displayed strong positive immunoreactivities against the recombinant N proteins, whereas normal sera gave negative immunoresponses to these proteins, indicating that the N protein of SARS-CoV is an antigenic protein. Furthermore, the epitope sites in the N protein were determined by competition experiments, in which the recombinant proteins or the synthesized peptides competed against the SARS-CoV proteins to bind to the antibodies raised in SARS sera. One epitope site located at the C-terminus was confirmed as the most antigenic region in this protein. A detailed screening of peptide with ELISA demonstrated that the amino sequence from Codons 371 to 407 was the epitope site at the C-terminus of the N protein. Understanding of the epitope sites could be very significant for developing an effective diagnostic approach to SARS.