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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| Format: | Article |
| Language: | English |
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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 |
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| Online Access: | https://www.academax.com/doi/10.3785/1008-9209.2005.02.0232 |
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| author | CHEN Hong-xun YAN Xing-cheng YING Jian-yue WU Jian-xiang ZHOU Ji-yong |
| author_facet | CHEN Hong-xun YAN Xing-cheng YING Jian-yue WU Jian-xiang ZHOU Ji-yong |
| author_sort | CHEN Hong-xun |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-09f0de4c63a5446ea5584dc741d98c93 |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2005-03-01 |
| publisher | Zhejiang University Press |
| record_format | Article |
| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-09f0de4c63a5446ea5584dc741d98c932025-08-20T03:34:22ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552005-03-013123223610.3785/1008-9209.2005.02.023210089209In vitro expression of SARS coronavirus (SARS-CoV) nucleocapsid protein and examination of the function of monoclonal antibody induced by the expressed proteinCHEN Hong-xunYAN Xing-chengYING Jian-yueWU Jian-xiangZHOU Ji-yongTo 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.https://www.academax.com/doi/10.3785/1008-9209.2005.02.0232SARScoronavirusnucleocapsid proteinmonoclonal antibody |
| spellingShingle | CHEN Hong-xun YAN Xing-cheng YING Jian-yue WU Jian-xiang ZHOU Ji-yong In vitro expression of SARS coronavirus (SARS-CoV) nucleocapsid protein and examination of the function of monoclonal antibody induced by the expressed protein 浙江大学学报. 农业与生命科学版 SARS coronavirus nucleocapsid protein monoclonal antibody |
| title | In vitro expression of SARS coronavirus (SARS-CoV) nucleocapsid protein and examination of the function of monoclonal antibody induced by the expressed protein |
| title_full | In vitro expression of SARS coronavirus (SARS-CoV) nucleocapsid protein and examination of the function of monoclonal antibody induced by the expressed protein |
| title_fullStr | In vitro expression of SARS coronavirus (SARS-CoV) nucleocapsid protein and examination of the function of monoclonal antibody induced by the expressed protein |
| title_full_unstemmed | In vitro expression of SARS coronavirus (SARS-CoV) nucleocapsid protein and examination of the function of monoclonal antibody induced by the expressed protein |
| title_short | In vitro expression of SARS coronavirus (SARS-CoV) nucleocapsid protein and examination of the function of monoclonal antibody induced by the expressed protein |
| title_sort | in vitro expression of sars coronavirus sars cov nucleocapsid protein and examination of the function of monoclonal antibody induced by the expressed protein |
| topic | SARS coronavirus nucleocapsid protein monoclonal antibody |
| url | https://www.academax.com/doi/10.3785/1008-9209.2005.02.0232 |
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