1.Production of specific antibodies against SARS-coronavirus nucleocapsid protein without cross reactivity with human coronaviruses 229E and OC43.
Hyun Kyoung LEE ; Byoung Hee LEE ; Seung Hyeok SEOK ; Min Won BAEK ; Hui Young LEE ; Dong Jae KIM ; Yi Rang NA ; Kyoung Jin NOH ; Sung Hoon PARK ; Dutta Noton KUMAR ; Hiroaki KARIWA ; Mina NAKAUCHI ; Suk Jin HEO ; Jae Hak PARK
Journal of Veterinary Science 2010;11(2):165-167
Severe acute respiratory syndrome (SARS) is a life-threatening disease for which accurate diagnosis is essential. Although many tools have been developed for the diagnosis of SARS, false-positive reactions in negative sera may occur because of cross-reactivity with other coronaviruses. We have raised polyclonal and monoclonal antibodies (Abs) using a recombinant form of the SARS virus nucleocapsid protein. Cross-reactivity of these anti-SARS Abs against human coronavirus (HCoV) 229E and HCoV OC43 were determined by Western blotting. The Abs produced reacted with recombinant SARS virus nucleocapsid protein, but not with HCoV 229E or HCoV OC43.
Antibodies, Viral/*immunology
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Blotting, Western
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Coronavirus 229E, Human/*immunology
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Coronavirus OC43, Human/*immunology
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Cross Reactions
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Humans
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Nucleocapsid Proteins/genetics/*immunology
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Recombinant Proteins/immunology
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SARS Virus/genetics/*immunology
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Severe Acute Respiratory Syndrome/diagnosis/*immunology
2.Antigenicity analysis of nucleocapsid proteins of 3 human coronaviruses SARS-CoV, 229E and OC43 with their monoclonal antibodies.
Li-wen QIU ; Ya-di WANG ; Zhi-yong LIAO ; Kun WEN ; Xiao-yan CHE
Journal of Southern Medical University 2006;26(3):290-293
OBJECTIVETo prepare and characterize monoclonal antibodies (mAbs) against the recombinant nucleocapsid (N) protein of 3 human coronaviruses SARS-CoV, 229E and OC43 and study the antigenic relationship between the 3 N proteins.
METHODSBALB/c mice were immunized with the recombinant N proteins of SARS-CoV, 229E and OC43 to obtain the mAbs by means of hybridoma. Screening and identification of the mAbs were performed using indirect enzyme-linked immunosorbent assay (ELISA), Western blotting and indirect immunofluorescence assay. Cross-reactivity between the N proteins of the 3 coronaviruses was analyzed with the prepared mAbs.
RESULTSThe mAbs against the recombinant N proteins of SARS-CoV, 229E and OC43 were obtained, which reacted specifically with the corresponding viral N protein as shown by indirect ELISA, Western blotting and indirect immunofluorescence assay. No cross-reactivity was found between the 3 N proteins.
CONCLUSIONThe prepared mAbs against the recombinant N proteins may provide valuable assistance in studying antigenic relationships of N proteins between the 3 human coronaviruses.
Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Coronavirus 229E, Human ; genetics ; immunology ; Coronavirus OC43, Human ; genetics ; immunology ; Cross Reactions ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Proteins ; immunology ; SARS Virus ; genetics ; immunology