Preparation and characterization of a novel monoclonal antibody specific to severe acute respiratory syndrome-coronavirus nucleocapsid protein.
Severe acute respiratory syndrome-coronavirus nucleocapsid (SARS-CoV N) protein has been found to be important to the processes related to viral pathogenesis, such as virus replication, interference of the cell process and modulation of host immune response; detection of the antigen has been used for the early diagnosis of infection. We have used recombinant N protein expressed in insect cells to generate 17 mAbs directed against this protein. We selected five mAbs that could be used in various diagnostic assays, and all of these mAbs recognized linear epitopes. Three IgG(2b) mAbs were recognized within the N-terminus of N protein, whereas the epitope of two IgG(1) mAbs localized within the C-terminus. These mAbs were found to have significant reactivity with both non-phosphorylated and phosphorylated N proteins, which resulted in high reactivity with native N protein in virus-infected cells; however, they did not show cross-reactivity with human coronavirus. Therefore, these results suggested that these mAbs would be useful in the development of various diagnostic kits and in future studies of SARS-CoV pathology.
Shin
Gu-Choul
GC
Division of Influenza and Respiratory Viruses, Center for Infectious Disease, National Institute of Health, Korea Center for Disease Control and Prevention, 5 Nokbun-dong, Eunpyung-gu, Seoul 122-701, Republic of Korea.
Chung
Yoon-Seok
YS
Kim
In-Soo
IS
Cho
Hae-Wol
HW
Kang
Chun
C
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity; D000956:Antigens, Viral / Q000276:immunology*; D002460:Cell Line; D000086462:Coronavirus Nucleocapsid Proteins; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D019590:Nucleocapsid Proteins / Q000276:immunology*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000086462:Coronavirus Nucleocapsid Proteins; D000939:Epitopes; D019590:Nucleocapsid Proteins