Development and characterization of an enterovirus 71 (EV71) virus-like particles (VLPs) vaccine produced in Pichia pastoris
Enterovirus 71 (EV71) is one of the major causative agents for hand, foot and mouth disease (HFMD) in children. Although there are three inactivated virus-based HFMD vaccines licensed in China, alternative approaches have been taken to produce an effective and safer vaccine that is easier to manufac...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2020-07-01
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| Series: | Human Vaccines & Immunotherapeutics |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/21645515.2019.1649554 |
| _version_ | 1797677354267443200 |
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| author | Zhijian Yang Fan Gao Xiaoliang Wang Likang Shi Zheng Zhou Yuanxiang Jiang Xinxing Ma Chao Zhang Chenliang Zhou Xianfang Zeng Ge Liu Jiang Fan Qunying Mao Li Shi |
| author_facet | Zhijian Yang Fan Gao Xiaoliang Wang Likang Shi Zheng Zhou Yuanxiang Jiang Xinxing Ma Chao Zhang Chenliang Zhou Xianfang Zeng Ge Liu Jiang Fan Qunying Mao Li Shi |
| author_sort | Zhijian Yang |
| collection | DOAJ |
| description | Enterovirus 71 (EV71) is one of the major causative agents for hand, foot and mouth disease (HFMD) in children. Although there are three inactivated virus-based HFMD vaccines licensed in China, alternative approaches have been taken to produce an effective and safer vaccine that is easier to manufacture in large scale. Among these, a virus-like particles (VLPs) based EV71 vaccine is under active development. For this purpose, an efficient methodology for the production of EV71-VLPs by recombinant technology is needed. We here report the construction and expression of the P1 and 3C genes of EV71 in Pichia pastoris for producing VLP-based EV71 vaccine antigen with a high yield and simple manufacturing process. Based on codon-optimized P1 and 3C genes, EV71-VLPs were efficiently expressed in Pichia pastoris system, and the expression level reached 270 mg/L. Biochemical and biophysical analyses showed that the produced EV71-VLPs consisted of processed VP0, VP1, and VP3 present as ~35nm spherical particles. The immune response as a function of EV71-VLPs and adjuvant dose ratio was investigated for vaccine development. Immunization with EV71-VLPs of 1–5 µg/dose and adjuvant of 225 µg/dose induced robust neutralizing antibody responses in mice and provided effective protection against lethal challenge in both maternally transferred antibody and passive transfer protection mouse models. Therefore, the yeast produced EV71-VLPs antigen is a promising candidate for the development of a vaccine against HFMD. |
| first_indexed | 2024-03-11T22:43:52Z |
| format | Article |
| id | doaj.art-8cb003d28c77409d8042192093133aa4 |
| institution | Directory Open Access Journal |
| issn | 2164-5515 2164-554X |
| language | English |
| last_indexed | 2024-03-11T22:43:52Z |
| publishDate | 2020-07-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Human Vaccines & Immunotherapeutics |
| spelling | doaj.art-8cb003d28c77409d8042192093133aa42023-09-22T08:45:32ZengTaylor & Francis GroupHuman Vaccines & Immunotherapeutics2164-55152164-554X2020-07-011671602161010.1080/21645515.2019.16495541649554Development and characterization of an enterovirus 71 (EV71) virus-like particles (VLPs) vaccine produced in Pichia pastorisZhijian Yang0Fan Gao1Xiaoliang Wang2Likang Shi3Zheng Zhou4Yuanxiang Jiang5Xinxing Ma6Chao Zhang7Chenliang Zhou8Xianfang Zeng9Ge Liu10Jiang Fan11Qunying Mao12Li Shi13Shanghai Zerun Biotechnology Co., LtdNational Institutes for Food and Drug Control (NIFDC)Shanghai Zerun Biotechnology Co., LtdShanghai Zerun Biotechnology Co., LtdShanghai Zerun Biotechnology Co., LtdShanghai Zerun Biotechnology Co., LtdShanghai Zerun Biotechnology Co., LtdShanghai Zerun Biotechnology Co., LtdShanghai Zerun Biotechnology Co., LtdShanghai Zerun Biotechnology Co., LtdShanghai Zerun Biotechnology Co., LtdShanghai Zerun Biotechnology Co., LtdNational Institutes for Food and Drug Control (NIFDC)Shanghai Zerun Biotechnology Co., LtdEnterovirus 71 (EV71) is one of the major causative agents for hand, foot and mouth disease (HFMD) in children. Although there are three inactivated virus-based HFMD vaccines licensed in China, alternative approaches have been taken to produce an effective and safer vaccine that is easier to manufacture in large scale. Among these, a virus-like particles (VLPs) based EV71 vaccine is under active development. For this purpose, an efficient methodology for the production of EV71-VLPs by recombinant technology is needed. We here report the construction and expression of the P1 and 3C genes of EV71 in Pichia pastoris for producing VLP-based EV71 vaccine antigen with a high yield and simple manufacturing process. Based on codon-optimized P1 and 3C genes, EV71-VLPs were efficiently expressed in Pichia pastoris system, and the expression level reached 270 mg/L. Biochemical and biophysical analyses showed that the produced EV71-VLPs consisted of processed VP0, VP1, and VP3 present as ~35nm spherical particles. The immune response as a function of EV71-VLPs and adjuvant dose ratio was investigated for vaccine development. Immunization with EV71-VLPs of 1–5 µg/dose and adjuvant of 225 µg/dose induced robust neutralizing antibody responses in mice and provided effective protection against lethal challenge in both maternally transferred antibody and passive transfer protection mouse models. Therefore, the yeast produced EV71-VLPs antigen is a promising candidate for the development of a vaccine against HFMD.http://dx.doi.org/10.1080/21645515.2019.1649554enterovirus 71hand foot and mouth diseasevaccineimmunogenicitypichia pastorisvirus-like particles |
| spellingShingle | Zhijian Yang Fan Gao Xiaoliang Wang Likang Shi Zheng Zhou Yuanxiang Jiang Xinxing Ma Chao Zhang Chenliang Zhou Xianfang Zeng Ge Liu Jiang Fan Qunying Mao Li Shi Development and characterization of an enterovirus 71 (EV71) virus-like particles (VLPs) vaccine produced in Pichia pastoris Human Vaccines & Immunotherapeutics enterovirus 71 hand foot and mouth disease vaccine immunogenicity pichia pastoris virus-like particles |
| title | Development and characterization of an enterovirus 71 (EV71) virus-like particles (VLPs) vaccine produced in Pichia pastoris |
| title_full | Development and characterization of an enterovirus 71 (EV71) virus-like particles (VLPs) vaccine produced in Pichia pastoris |
| title_fullStr | Development and characterization of an enterovirus 71 (EV71) virus-like particles (VLPs) vaccine produced in Pichia pastoris |
| title_full_unstemmed | Development and characterization of an enterovirus 71 (EV71) virus-like particles (VLPs) vaccine produced in Pichia pastoris |
| title_short | Development and characterization of an enterovirus 71 (EV71) virus-like particles (VLPs) vaccine produced in Pichia pastoris |
| title_sort | development and characterization of an enterovirus 71 ev71 virus like particles vlps vaccine produced in pichia pastoris |
| topic | enterovirus 71 hand foot and mouth disease vaccine immunogenicity pichia pastoris virus-like particles |
| url | http://dx.doi.org/10.1080/21645515.2019.1649554 |
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