@ARTICLE{Wang_Y.B._Expression_2023,
 author={Wang, Y.B. and Wang, S.W. and Jin, Q.Y. and Chen, L.P. and Zhang, F.Q. and Shi, J.J. and Yin, Y. and Fan, Z.X. and Liu, X.Y. and Wang, L.P. and Li, P.},
 volume={vol. 26},
 number={No 4},
 journal={Polish Journal of Veterinary Sciences},
 pages={571-579},
 howpublished={online},
 year={2023},
 publisher={Polish Academy of Sciences Committee of Veterinary Sciences},
 publisher={University of Warmia and Mazury in Olsztyn},
 abstract={Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to be a major public health concern. Nucleocapsid (N) protein is the most abundant structural protein on SARS-CoV-2 virions and induces the production of antibodies at the early stage of infection. Large-scale preparation of N protein is essential for the development of immunoassays to detect antibodies to SARS-CoV-2 and the control of virus transmission. In this study, expression of water-soluble N protein was achieved through inducing protein expression at 25°C with 0.5 mM IPTG for 12 h. Western blot and ELISA showed that recombinant N protein could be recognized by sera collected from subjects immunized with Sinovac inactivated SARS-CoV-2 vaccine. Four monoclonal antibodies namely 2B1B1, 4D3A3, 5G1F8, and 7C6F5 were produced using hybridoma technology. Titers of all four monoclonal antibodies in ELISA reached more than 1.28×10 6.0. Moreover, all monoclonal antibodies could react specifically with N protein expressed by transfection of pcDNA3.1-N into BHK-21 cells in IPMA and IFA. These results indicated that water-soluble N protein retained high immunogenicity and possessed the same epitopes as that of native N protein on virions. In addition, the preparation of water-soluble N protein and its monoclonal antibodies laid the basis for the development of immunoassays for COVID-19 detection.},
 type={Article},
 title={Expression of water-soluble nucleocapsid protein of SARS-CoV-2 and analysis of its immunogenicity},
 URL={http://journals.pan.pl/Content/129581/3%20_%20Dik.pdf},
 doi={10.24425/pjvs.2023.148277},
 keywords={SARS-CoV-2, optimization of expression conditions, water-soluble nucleocapsid protein, immunogenicity analysis},
}