[Preparation of specific monoclonal antibody against nucleocapsid protein of SARS coronavirus].
OBJECTIVE
To express nucleocapsid(N) protein of SARS coronavirus and produce monoclonal antibody(mAb) to N protein.


METHODS
N protein gene was amplified by RT-PCR. After being confirmed by DNA sequencing, the gene was subcloned into prokaryotic expression vector. N protein expressed in E.coli was recovered from SDS-PAGE gel and served as immunogen in the preparation of the mAb.


RESULTS
DNA sequencing confirmed that the amplified fragment was N protein gene. SDS-PAGE analysis showed that the M(r) of the expressed protein was approximately 43 kd. This expressed protein could be further confirmed in Western blot by using serum from a convalescent SARS patient as primary antibody. Western blot analysis proved that three mAbs obtained could react specifically to the recombinant N protein.


CONCLUSIONS
The prepared recombinant N protein and mAbs against N protein lay the foundation for further development of early diagnosis assays for SARS coronavirus infection.
Wang
Ping
P
Department of Microbiology, Fourth Military Medical University, Xi'an 710032, China.
Huang
Qing-Sheng
QS
Xue
Xiao-Ping
XP
Lei
Ying-Feng
YF
Wang
Hai-Tao
HT
Ren
Jun-Ping
JP
Ding
Tian-Bing
TB
Xu
Zhi-Kai
ZK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D051379:Mice; D019590:Nucleocapsid Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*; D017422:Sequence Analysis, DNA; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis / Q000276:immunology
D000911:Antibodies, Monoclonal; D019590:Nucleocapsid Proteins