Identification and epitope mapping of swine acute diarrhea syndrome coronavirus accessory protein NS7a via monoclonal antibodies
Swine acute diarrhea syndrome coronavirus (SADS-CoV) is an emerging swine enteric coronavirus that causes vomiting, severe diarrhea, dehydration and death in suckling piglets. NS7a is putative accessory protein that is predicted to be encoded by SADS-CoV, but still to be confirmed experimentally. In...
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Elsevier
2022-05-01
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| Series: | Virus Research |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0168170222000703 |
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| author | Yibin Qin Tingshuai Feng Hongyan Shi Jiyu Zhang Liaoyuan Zhang Shufeng Feng Jianfei Chen Ying He Xin Zhang Zhongwei Chen Jianbo Liu Dakai Liu Da Shi Li Feng |
| author_facet | Yibin Qin Tingshuai Feng Hongyan Shi Jiyu Zhang Liaoyuan Zhang Shufeng Feng Jianfei Chen Ying He Xin Zhang Zhongwei Chen Jianbo Liu Dakai Liu Da Shi Li Feng |
| author_sort | Yibin Qin |
| collection | DOAJ |
| description | Swine acute diarrhea syndrome coronavirus (SADS-CoV) is an emerging swine enteric coronavirus that causes vomiting, severe diarrhea, dehydration and death in suckling piglets. NS7a is putative accessory protein that is predicted to be encoded by SADS-CoV, but still to be confirmed experimentally. In the present study, recombinant NS7a protein was expressed in a prokaryotic expression system and used as an antigen to prepare monoclonal antibodies (mAbs) specific to NS7a protein. We obtained two anti-NS7a mAbs, termed AH5 and EH3, that were shown by western blotting to react with the natural NS7a protein in Vero E6 cells infected with SADS-CoV. Using the produced mAbs, we observed by confocal microscopy that NS7a protein was expressed in the cytoplasm. Further studies revealed that the motif 31VNTWQEFA38 was the minimal unit of the linear B-cell epitope recognized by mAb AH5, and the motif 82FDLFERF88 was the minimal unit of the linear B-cell epitope recognized by mAb EH3. Alignment of amino acids showed that these two epitopes were highly conserved among different SADS-CoV strains and SADS-related coronaviruses from bats, but with one substitution in these two motifs in bat coronavirus HKU2. In summary, we generated and characterized two mAbs against SADS-CoV NS7a protein, and demonstrated NS7a expression in SADS-CoV-infected cells for the first time. |
| first_indexed | 2024-03-08T17:07:07Z |
| format | Article |
| id | doaj.art-a0de5e912c9d45438f9c00a6a43f207d |
| institution | Directory Open Access Journal |
| issn | 1872-7492 |
| language | English |
| last_indexed | 2024-03-08T17:07:07Z |
| publishDate | 2022-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Virus Research |
| spelling | doaj.art-a0de5e912c9d45438f9c00a6a43f207d2024-01-04T04:35:59ZengElsevierVirus Research1872-74922022-05-01313198742Identification and epitope mapping of swine acute diarrhea syndrome coronavirus accessory protein NS7a via monoclonal antibodiesYibin Qin0Tingshuai Feng1Hongyan Shi2Jiyu Zhang3Liaoyuan Zhang4Shufeng Feng5Jianfei Chen6Ying He7Xin Zhang8Zhongwei Chen9Jianbo Liu10Dakai Liu11Da Shi12Li Feng13State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, China; Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Xixiangtang District, Youai North Road 51, Nanning, 530001, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Xixiangtang District, Youai North Road 51, Nanning, 530001, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research Institute, Xixiangtang District, Youai North Road 51, Nanning, 530001, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xiangfang District, Haping Road 678, Harbin, 150069, China; Corresponding author at: Harbin Veterinary Research Institute, CAAS, 678 Haping Road Xiangfang District, Harbin 150069, China.Swine acute diarrhea syndrome coronavirus (SADS-CoV) is an emerging swine enteric coronavirus that causes vomiting, severe diarrhea, dehydration and death in suckling piglets. NS7a is putative accessory protein that is predicted to be encoded by SADS-CoV, but still to be confirmed experimentally. In the present study, recombinant NS7a protein was expressed in a prokaryotic expression system and used as an antigen to prepare monoclonal antibodies (mAbs) specific to NS7a protein. We obtained two anti-NS7a mAbs, termed AH5 and EH3, that were shown by western blotting to react with the natural NS7a protein in Vero E6 cells infected with SADS-CoV. Using the produced mAbs, we observed by confocal microscopy that NS7a protein was expressed in the cytoplasm. Further studies revealed that the motif 31VNTWQEFA38 was the minimal unit of the linear B-cell epitope recognized by mAb AH5, and the motif 82FDLFERF88 was the minimal unit of the linear B-cell epitope recognized by mAb EH3. Alignment of amino acids showed that these two epitopes were highly conserved among different SADS-CoV strains and SADS-related coronaviruses from bats, but with one substitution in these two motifs in bat coronavirus HKU2. In summary, we generated and characterized two mAbs against SADS-CoV NS7a protein, and demonstrated NS7a expression in SADS-CoV-infected cells for the first time.http://www.sciencedirect.com/science/article/pii/S0168170222000703Swine acute diarrhea syndrome coronavirusNS7a proteinMonoclonal antibodyEpitope mapping |
| spellingShingle | Yibin Qin Tingshuai Feng Hongyan Shi Jiyu Zhang Liaoyuan Zhang Shufeng Feng Jianfei Chen Ying He Xin Zhang Zhongwei Chen Jianbo Liu Dakai Liu Da Shi Li Feng Identification and epitope mapping of swine acute diarrhea syndrome coronavirus accessory protein NS7a via monoclonal antibodies Virus Research Swine acute diarrhea syndrome coronavirus NS7a protein Monoclonal antibody Epitope mapping |
| title | Identification and epitope mapping of swine acute diarrhea syndrome coronavirus accessory protein NS7a via monoclonal antibodies |
| title_full | Identification and epitope mapping of swine acute diarrhea syndrome coronavirus accessory protein NS7a via monoclonal antibodies |
| title_fullStr | Identification and epitope mapping of swine acute diarrhea syndrome coronavirus accessory protein NS7a via monoclonal antibodies |
| title_full_unstemmed | Identification and epitope mapping of swine acute diarrhea syndrome coronavirus accessory protein NS7a via monoclonal antibodies |
| title_short | Identification and epitope mapping of swine acute diarrhea syndrome coronavirus accessory protein NS7a via monoclonal antibodies |
| title_sort | identification and epitope mapping of swine acute diarrhea syndrome coronavirus accessory protein ns7a via monoclonal antibodies |
| topic | Swine acute diarrhea syndrome coronavirus NS7a protein Monoclonal antibody Epitope mapping |
| url | http://www.sciencedirect.com/science/article/pii/S0168170222000703 |
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