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Preparation of specific monoclonal antibody against nucleocapsid protein of SARS coronavirus



Preparation of specific monoclonal antibody against nucleocapsid protein of SARS coronavirus



Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 21(1): 50-52



To express nucleocapsid(N) protein of SARS coronavirus and produce monoclonal antibody(mAb) to N protein. 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. 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. The prepared recombinant N protein and mAbs against N protein lay the foundation for further development of early diagnosis assays for SARS coronavirus infection.

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Accession: 049990455

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PMID: 15629083


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