In vitro expression of SARS coronavirus (SARS-CoV) nucleocapsid protein and examination of the function of monoclonal antibody induced by the expressed protein
To efficiently express SARS coronavirus (SARS-CoV) nucleocapsid (N) protein in vitro and produce monoclonal antibody (mAb) against SARS-CoV N protein, the recombinant clone PGEX-4T/N was induced with IPTG to express target protein and the fusion protein (GST-N) reacted specifically with anti-GST pol...
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| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2005-03-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/1008-9209.2005.02.0232 |
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| Summary: | To efficiently express SARS coronavirus (SARS-CoV) nucleocapsid (N) protein in vitro and produce monoclonal antibody (mAb) against SARS-CoV N protein, the recombinant clone PGEX-4T/N was induced with IPTG to express target protein and the fusion protein (GST-N) reacted specifically with anti-GST polyclonal antibody. The expressed recombinant protein was purified by affinity chromatography and gel extraction, where the purified protein concentrations were 0.85 mg/mL and 0.433 mg/mL respectively. Four hybridoma cell lines secreting monoclonal antibody (mAb) against SARS-CoV N protein were obtained and named as 3E5, 6B9, 4C2, 4B7. Subtype analysis revealed that mAbs 3E5, 4C2 and 4B7 against SARS-CoV N protein belong to IgG<sub>1</sub> while 6B9 is allied to IgG<sub>2a</sub>. Western blot analysis showed that the four mAbs could react with SARS-CoV N protein but not with N protein of infectious bronchitis virus. |
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| ISSN: | 1008-9209 2097-5155 |