文献类型： 外文期刊

作者： Wang, Y. B. ¹ ; Wang, S. W. ² ; Jin, Q. Y. ³ ; Chen, L. P. ⁴ ; Zhang, F. Q. ¹ ; Shi, J. J. ¹ ; Yin, Y. ⁵ ; Fan, Z. X. ¹ ; Lin, X. Y. ⁵ ; Wang, L. P. ⁶ ; Li, P. ⁶ ;

作者机构： 1.Xinxiang Med Univ, Sch Publ Hlth, Xinxiang 453003, Peoples R China

2.Xinxiang Med Univ, Sch Basic Med Sci, Xinxiang 453003, Peoples R China

3.Henan Acad Agr Sci, Henan Prov Key Lab Anim Immunol, Zhengzhou 450002, Peoples R China

4.Gushi Cty Ctr Anim Dis Control & Prevent, Xinyang 465200, Peoples R China

5.Xinxiang Med Univ, Mingde Coll, Xinxiang 453003, Peoples R China

6.Xinxiang Univ, Sch Biol Engn, Xinxiang 453003, Peoples R China

关键词： SARS-CoV-2; optimization of expression conditions; water-soluble nucleocapsid protein; immunogenicity analysis

期刊名称：POLISH JOURNAL OF VETERINARY SCIENCES （ 影响因子：0.8； 五年影响因子：0.9 ）

ISSN： 1505-1773

年卷期： 2023 年 26 卷 4 期

页码：

收录情况： SCI

摘要： Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome corona -virus 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 degrees 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 1.28x106.0. 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 immuno-genicity 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.