Screening of strategies for HA protein vaccine for influenza virus H1N1
Objective To screen and establish the optimal expression strategy of influenza virus H1N1 A/Nebraska/14/2019 HA, and lay a foundation for the development of protein vaccines. Methods The recombinant baculovirus of HA full-length, different C-terminal mutants of the extra cellular regions and co-ex...
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| Format: | Article |
| Language: | zho |
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Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.
2022-12-01
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| Series: | Jichu yixue yu linchuang |
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| Online Access: | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-12-1885.pdf |
| _version_ | 1827390596300406784 |
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| author | ZHENG Ning-chen, YANG Jiao-jiao, ZHANG Ting, WANG Zhi-rong, XU Xue-mei |
| author_facet | ZHENG Ning-chen, YANG Jiao-jiao, ZHANG Ting, WANG Zhi-rong, XU Xue-mei |
| author_sort | ZHENG Ning-chen, YANG Jiao-jiao, ZHANG Ting, WANG Zhi-rong, XU Xue-mei |
| collection | DOAJ |
| description | Objective To screen and establish the optimal expression strategy of influenza virus H1N1 A/Nebraska/14/2019 HA, and lay a foundation for the development of protein vaccines. Methods The recombinant baculovirus of HA full-length, different C-terminal mutants of the extra cellular regions and co-expressing HA/M1 were constructed respectively. After 80 hours of infection in insect cells, the culture supernatant was collected, and the expression of recombinant antigen was identified by Western blot. Hemagglutination test indirectly analyzes its immunogenic. Then, HA/M1 protein was expressed in large quantities, and the physicochemical properties, hemagglutination activity and stability were analyzed after Core700 purification. Results The two C-terminal mutants of the extra cellular region of HA were all expressed in the form of trimers, but the expression supernatants had no hemagglutination activity; the full-length HA trimers had good hemagglutination activity, but they were membrane proteins thus were difficult to be purified. M1/HA co-expression supernatant also had hemagglutination activity. Electron microscopy of expression product purified by Core700 showed virus-like particles (VLPs) of 80 nm~150 nm. The hemagglutination titer of HA-M1-VLP reached 26 and was stable for three months at 4 ℃ as shown by light scattering. Conclusions HA-M1-VLP is easy to express and to be purified. The product has good stability, and hemagglutination activity, so can be used for the research of influenza virus protein vaccine. |
| first_indexed | 2024-03-08T16:55:02Z |
| format | Article |
| id | doaj.art-842586190af04957963fe52b03154e51 |
| institution | Directory Open Access Journal |
| issn | 1001-6325 |
| language | zho |
| last_indexed | 2024-03-08T16:55:02Z |
| publishDate | 2022-12-01 |
| publisher | Institute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College. |
| record_format | Article |
| series | Jichu yixue yu linchuang |
| spelling | doaj.art-842586190af04957963fe52b03154e512024-01-05T02:36:47ZzhoInstitute of Basic Medical Sciences and Peking Union Medical College Hospital, Chinese Academy of Medical Sciences / Peking Union Medical College.Jichu yixue yu linchuang1001-63252022-12-0142121885189010.16352/j.issn.1001-6325.2022.12.1885Screening of strategies for HA protein vaccine for influenza virus H1N1ZHENG Ning-chen, YANG Jiao-jiao, ZHANG Ting, WANG Zhi-rong, XU Xue-mei0Department of Biophysics and Structure Biology,Institute of Basic Medical Sciences CAMS,School of Basic Medicine PUMC, Beijing 100005,ChinaObjective To screen and establish the optimal expression strategy of influenza virus H1N1 A/Nebraska/14/2019 HA, and lay a foundation for the development of protein vaccines. Methods The recombinant baculovirus of HA full-length, different C-terminal mutants of the extra cellular regions and co-expressing HA/M1 were constructed respectively. After 80 hours of infection in insect cells, the culture supernatant was collected, and the expression of recombinant antigen was identified by Western blot. Hemagglutination test indirectly analyzes its immunogenic. Then, HA/M1 protein was expressed in large quantities, and the physicochemical properties, hemagglutination activity and stability were analyzed after Core700 purification. Results The two C-terminal mutants of the extra cellular region of HA were all expressed in the form of trimers, but the expression supernatants had no hemagglutination activity; the full-length HA trimers had good hemagglutination activity, but they were membrane proteins thus were difficult to be purified. M1/HA co-expression supernatant also had hemagglutination activity. Electron microscopy of expression product purified by Core700 showed virus-like particles (VLPs) of 80 nm~150 nm. The hemagglutination titer of HA-M1-VLP reached 26 and was stable for three months at 4 ℃ as shown by light scattering. Conclusions HA-M1-VLP is easy to express and to be purified. The product has good stability, and hemagglutination activity, so can be used for the research of influenza virus protein vaccine.http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-12-1885.pdfh1n1|hemagglutinin (ha)|matrix protein(m1)|virus-like particle (vlp) |
| spellingShingle | ZHENG Ning-chen, YANG Jiao-jiao, ZHANG Ting, WANG Zhi-rong, XU Xue-mei Screening of strategies for HA protein vaccine for influenza virus H1N1 Jichu yixue yu linchuang h1n1|hemagglutinin (ha)|matrix protein(m1)|virus-like particle (vlp) |
| title | Screening of strategies for HA protein vaccine for influenza virus H1N1 |
| title_full | Screening of strategies for HA protein vaccine for influenza virus H1N1 |
| title_fullStr | Screening of strategies for HA protein vaccine for influenza virus H1N1 |
| title_full_unstemmed | Screening of strategies for HA protein vaccine for influenza virus H1N1 |
| title_short | Screening of strategies for HA protein vaccine for influenza virus H1N1 |
| title_sort | screening of strategies for ha protein vaccine for influenza virus h1n1 |
| topic | h1n1|hemagglutinin (ha)|matrix protein(m1)|virus-like particle (vlp) |
| url | http://journal11.magtechjournal.com/Jwk_jcyxylc/fileup/1001-6325/PDF/1001-6325-2022-42-12-1885.pdf |
| work_keys_str_mv | AT zhengningchenyangjiaojiaozhangtingwangzhirongxuxuemei screeningofstrategiesforhaproteinvaccineforinfluenzavirush1n1 |