Production and identification of monoclonal antibodies and development of a sandwich ELISA for detection of the H3-subtype avian influenza virus antigen
Abstract The H3 subtype of avian influenza virus (AIV) is widespread in avian species and is frequently isolated in surveillance projects; thus, we have developed a more effective diagnostic approach of a monoclonal antibody (mAb)-based sandwich ELISA for the H3 AIV detection. First, we have produce...
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| Format: | Article |
| Language: | English |
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SpringerOpen
2020-03-01
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| Series: | AMB Express |
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| Online Access: | http://link.springer.com/article/10.1186/s13568-020-00988-7 |
| _version_ | 1811327297869840384 |
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| author | Sisi Luo Xianwen Deng Zhixun Xie Jiaoling Huang Minxiu Zhang Meng Li Liji Xie Dan Li Qing Fan Sheng Wang Tingting Zeng Yanfang Zhang Zhiqin Xie |
| author_facet | Sisi Luo Xianwen Deng Zhixun Xie Jiaoling Huang Minxiu Zhang Meng Li Liji Xie Dan Li Qing Fan Sheng Wang Tingting Zeng Yanfang Zhang Zhiqin Xie |
| author_sort | Sisi Luo |
| collection | DOAJ |
| description | Abstract The H3 subtype of avian influenza virus (AIV) is widespread in avian species and is frequently isolated in surveillance projects; thus, we have developed a more effective diagnostic approach of a monoclonal antibody (mAb)-based sandwich ELISA for the H3 AIV detection. First, we have produced the essential reagent of mAb against AIV H3 strains with the development of an mAb-Mouse immunization with a purified H3-subtype AIV strain and cell fusion to generate hybridoma cells. These cells were screened with hemagglutination inhibition (HI) tests, and optimal cells were subcloned. We chose a hybridoma cell line that steadily secreted a specific H3-subtype AIV mAb, designated 9F12, that belongs to the IgG1 subclass and has a K-type light chain. 9F12 was shown to bind specifically to the H3-subtype AIV antigen by both immunofluorescence assay and Western blot analysis. Finally, a 9F12-based sandwich ELISA was successfully developed and used to specifically test for this antigen. The sandwich ELISA conditions were optimized, and the specificity and sensitivity were validated. The results for clinical sample detection were consistent with viral isolation. Consequently, the 9F12-based sandwich ELISA is a specific, sensitive, robust, rapid and versatile diagnostic tool for H3-subtype AIV and provides a promising strategy for effective influenza virus prevention and control. |
| first_indexed | 2024-04-13T15:05:17Z |
| format | Article |
| id | doaj.art-37b34bb5403f499e8cc5c472e1360566 |
| institution | Directory Open Access Journal |
| issn | 2191-0855 |
| language | English |
| last_indexed | 2024-04-13T15:05:17Z |
| publishDate | 2020-03-01 |
| publisher | SpringerOpen |
| record_format | Article |
| series | AMB Express |
| spelling | doaj.art-37b34bb5403f499e8cc5c472e13605662022-12-22T02:42:11ZengSpringerOpenAMB Express2191-08552020-03-011011910.1186/s13568-020-00988-7Production and identification of monoclonal antibodies and development of a sandwich ELISA for detection of the H3-subtype avian influenza virus antigenSisi Luo0Xianwen Deng1Zhixun Xie2Jiaoling Huang3Minxiu Zhang4Meng Li5Liji Xie6Dan Li7Qing Fan8Sheng Wang9Tingting Zeng10Yanfang Zhang11Zhiqin Xie12Guangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteGuangxi Key Laboratory of Veterinary Biotechnology, Guangxi Veterinary Research InstituteAbstract The H3 subtype of avian influenza virus (AIV) is widespread in avian species and is frequently isolated in surveillance projects; thus, we have developed a more effective diagnostic approach of a monoclonal antibody (mAb)-based sandwich ELISA for the H3 AIV detection. First, we have produced the essential reagent of mAb against AIV H3 strains with the development of an mAb-Mouse immunization with a purified H3-subtype AIV strain and cell fusion to generate hybridoma cells. These cells were screened with hemagglutination inhibition (HI) tests, and optimal cells were subcloned. We chose a hybridoma cell line that steadily secreted a specific H3-subtype AIV mAb, designated 9F12, that belongs to the IgG1 subclass and has a K-type light chain. 9F12 was shown to bind specifically to the H3-subtype AIV antigen by both immunofluorescence assay and Western blot analysis. Finally, a 9F12-based sandwich ELISA was successfully developed and used to specifically test for this antigen. The sandwich ELISA conditions were optimized, and the specificity and sensitivity were validated. The results for clinical sample detection were consistent with viral isolation. Consequently, the 9F12-based sandwich ELISA is a specific, sensitive, robust, rapid and versatile diagnostic tool for H3-subtype AIV and provides a promising strategy for effective influenza virus prevention and control.http://link.springer.com/article/10.1186/s13568-020-00988-7H3 subtypeInfluenzaMonoclonal antibodyELISA |
| spellingShingle | Sisi Luo Xianwen Deng Zhixun Xie Jiaoling Huang Minxiu Zhang Meng Li Liji Xie Dan Li Qing Fan Sheng Wang Tingting Zeng Yanfang Zhang Zhiqin Xie Production and identification of monoclonal antibodies and development of a sandwich ELISA for detection of the H3-subtype avian influenza virus antigen AMB Express H3 subtype Influenza Monoclonal antibody ELISA |
| title | Production and identification of monoclonal antibodies and development of a sandwich ELISA for detection of the H3-subtype avian influenza virus antigen |
| title_full | Production and identification of monoclonal antibodies and development of a sandwich ELISA for detection of the H3-subtype avian influenza virus antigen |
| title_fullStr | Production and identification of monoclonal antibodies and development of a sandwich ELISA for detection of the H3-subtype avian influenza virus antigen |
| title_full_unstemmed | Production and identification of monoclonal antibodies and development of a sandwich ELISA for detection of the H3-subtype avian influenza virus antigen |
| title_short | Production and identification of monoclonal antibodies and development of a sandwich ELISA for detection of the H3-subtype avian influenza virus antigen |
| title_sort | production and identification of monoclonal antibodies and development of a sandwich elisa for detection of the h3 subtype avian influenza virus antigen |
| topic | H3 subtype Influenza Monoclonal antibody ELISA |
| url | http://link.springer.com/article/10.1186/s13568-020-00988-7 |
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