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Journal of Clinical Microbiology, November 2004, p. 5309-5314, Vol. 42, No. 11
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.11.5309-5314.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Mapping of Antigenic Sites on the Nucleocapsid Protein of the Severe Acute Respiratory Syndrome Coronavirus

Yuxian He,1 Yusen Zhou,2* Hao Wu,3 Zhihua Kou,2 Shuwen Liu,1 and Shibo Jiang1*

Lindsley F. Kimball Research Institute, New York Blood Center, New York, New York,1 Beijing Institute of Microbiology and Epidemiology,2 YouAn Hospital, Capital University of Medical Sciences, Beijing, China3

Received 9 June 2004/ Returned for modification 18 July 2004/ Accepted 27 July 2004

Antigenic sites on the nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) were mapped by Pepscan analysis with overlapping peptides that span the N protein sequence. Two major immunodominant epitopes located in the C-terminal region (amino acids [aa] 362 to 412) and middle region (aa 153 to 178) reacted with more than 75% of sera from SARS patients. Several minor immunodominant epitopes were reactive with about 50% of the SARS sera. Antisera from mice immunized with inactivated SARS-CoV recognized the two major immunodominant epitopes and one antigenic site located adjacent to the N-terminal region (aa 76 to 101), which did not react with the sera from SARS patients. Several monoclonal antibodies against SARS-CoV bound to the N- or C-terminal antigenic sites. These results suggest that the above antigenic sites on the N protein are important in eliciting humoral immune response against SARS-CoV in humans and animals and can be used as antigens for developing diagnostic tests.

* Corresponding author. Mailing address for Shibo Jiang: Lindsley F. Kimball Research Institute, New York Blood Center, 310 East 67th St., New York, NY 10021. Phone: (212) 570-3058. Fax: (212) 570-3099. E-mail: SJiang{at}NYBloodcenter.org. Mailing address for Yusen Zhou: Beijing Institute of Microbiology and Epidemiology, Beijing, China. Phone: 86-10-66948580. Fax: 86-10-66948490. E-mail: zhouy501{at}yahoo.com.

Journal of Clinical Microbiology, November 2004, p. 5309-5314, Vol. 42, No. 11
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.11.5309-5314.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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