Production and Immunogenicity Assessment of LTp50: An <i>Escherichia coli</i>-Made Chimeric Antigen Targeting S1- and S2-Epitopes from the SARS-CoV-2/BA.5 Spike Protein
Subunit vaccines stand as a leading approach to expanding the current portfolio of vaccines to fight against COVID-19, seeking not only to lower costs but to achieve long-term immunity against variants of concern and have the main attributes that could overcome the limitations of the current vaccine...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-02-01
|
| Series: | Pharmaceuticals |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1424-8247/17/3/302 |
| _version_ | 1827305314946383872 |
|---|---|
| author | Alejandra Wong-Arce Omar Gonzalez-Ortega Andrea Romero-Maldonado Arleth Miranda-López Mariano García-Soto Susan Farfán-Castro Lourdes Betancourt-Mendiola Samaporn Teeravechyan Kanjana Srisutthisamphan Mauricio Comas-García Karla I. Solís Andrade Sergio Rosales-Mendoza |
| author_facet | Alejandra Wong-Arce Omar Gonzalez-Ortega Andrea Romero-Maldonado Arleth Miranda-López Mariano García-Soto Susan Farfán-Castro Lourdes Betancourt-Mendiola Samaporn Teeravechyan Kanjana Srisutthisamphan Mauricio Comas-García Karla I. Solís Andrade Sergio Rosales-Mendoza |
| author_sort | Alejandra Wong-Arce |
| collection | DOAJ |
| description | Subunit vaccines stand as a leading approach to expanding the current portfolio of vaccines to fight against COVID-19, seeking not only to lower costs but to achieve long-term immunity against variants of concern and have the main attributes that could overcome the limitations of the current vaccines. Herein a chimeric protein targeting S1 and S2 epitopes, called LTp50, was designed as a convenient approach to induce humoral responses against SARS-CoV-2. LTp50 was produced in recombinant <i>Escherichia coli</i> using a conventional pET vector, recovering the expected antigen in the insoluble fraction. LTp50 was purified by chromatography (purity > 90%). The solubilization and refolding stages helped to obtain a stable protein amenable for vaccine formulation. LTp50 was adsorbed onto alum, resulting in a stable formulation whose immunogenic properties were assessed in BALB/c mice. Significant humoral responses against the S protein (BA.5 variant) were detected in mice subjected to three subcutaneous doses (10 µg) of the LTp50/alum formulation. This study opens the path for the vaccine formulation optimization using additional adjuvants to advance in the development of a highly effective anti-COVID-19 vaccine directed against the antigenic regions of the S protein, which are less prone to mutations. |
| first_indexed | 2024-04-24T17:56:08Z |
| format | Article |
| id | doaj.art-748b7351c59043fd998c4ca112a6f8a4 |
| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-04-24T17:56:08Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceuticals |
| spelling | doaj.art-748b7351c59043fd998c4ca112a6f8a42024-03-27T13:59:14ZengMDPI AGPharmaceuticals1424-82472024-02-0117330210.3390/ph17030302Production and Immunogenicity Assessment of LTp50: An <i>Escherichia coli</i>-Made Chimeric Antigen Targeting S1- and S2-Epitopes from the SARS-CoV-2/BA.5 Spike ProteinAlejandra Wong-Arce0Omar Gonzalez-Ortega1Andrea Romero-Maldonado2Arleth Miranda-López3Mariano García-Soto4Susan Farfán-Castro5Lourdes Betancourt-Mendiola6Samaporn Teeravechyan7Kanjana Srisutthisamphan8Mauricio Comas-García9Karla I. Solís Andrade10Sergio Rosales-Mendoza11Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, San Luis Potosí 78210, MexicoFacultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, San Luis Potosí 78210, MexicoFacultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, San Luis Potosí 78210, MexicoFacultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, San Luis Potosí 78210, MexicoFacultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, San Luis Potosí 78210, MexicoFacultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, San Luis Potosí 78210, MexicoFacultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, San Luis Potosí 78210, MexicoVirology and Cell Technology Laboratory, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park Phaholyothin Rd, Klong 1, Klong Luang, Pathumthani 12120, ThailandVirology and Cell Technology Laboratory, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park Phaholyothin Rd, Klong 1, Klong Luang, Pathumthani 12120, ThailandSección de Microscopía de Alta Resolución, Centro de Investigación en Ciencias de la Salud y Biomedicina, Universidad Autónoma de San Luis Potosí, Av. Sierra Leona 550, Lomas 2a Sección, San Luis Potosí 78210, MexicoFacultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, San Luis Potosí 78210, MexicoFacultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, San Luis Potosí 78210, MexicoSubunit vaccines stand as a leading approach to expanding the current portfolio of vaccines to fight against COVID-19, seeking not only to lower costs but to achieve long-term immunity against variants of concern and have the main attributes that could overcome the limitations of the current vaccines. Herein a chimeric protein targeting S1 and S2 epitopes, called LTp50, was designed as a convenient approach to induce humoral responses against SARS-CoV-2. LTp50 was produced in recombinant <i>Escherichia coli</i> using a conventional pET vector, recovering the expected antigen in the insoluble fraction. LTp50 was purified by chromatography (purity > 90%). The solubilization and refolding stages helped to obtain a stable protein amenable for vaccine formulation. LTp50 was adsorbed onto alum, resulting in a stable formulation whose immunogenic properties were assessed in BALB/c mice. Significant humoral responses against the S protein (BA.5 variant) were detected in mice subjected to three subcutaneous doses (10 µg) of the LTp50/alum formulation. This study opens the path for the vaccine formulation optimization using additional adjuvants to advance in the development of a highly effective anti-COVID-19 vaccine directed against the antigenic regions of the S protein, which are less prone to mutations.https://www.mdpi.com/1424-8247/17/3/302linear epitopesbuilt-in adjuvanthumoral responsechimeric antigenCOVID-19 |
| spellingShingle | Alejandra Wong-Arce Omar Gonzalez-Ortega Andrea Romero-Maldonado Arleth Miranda-López Mariano García-Soto Susan Farfán-Castro Lourdes Betancourt-Mendiola Samaporn Teeravechyan Kanjana Srisutthisamphan Mauricio Comas-García Karla I. Solís Andrade Sergio Rosales-Mendoza Production and Immunogenicity Assessment of LTp50: An <i>Escherichia coli</i>-Made Chimeric Antigen Targeting S1- and S2-Epitopes from the SARS-CoV-2/BA.5 Spike Protein Pharmaceuticals linear epitopes built-in adjuvant humoral response chimeric antigen COVID-19 |
| title | Production and Immunogenicity Assessment of LTp50: An <i>Escherichia coli</i>-Made Chimeric Antigen Targeting S1- and S2-Epitopes from the SARS-CoV-2/BA.5 Spike Protein |
| title_full | Production and Immunogenicity Assessment of LTp50: An <i>Escherichia coli</i>-Made Chimeric Antigen Targeting S1- and S2-Epitopes from the SARS-CoV-2/BA.5 Spike Protein |
| title_fullStr | Production and Immunogenicity Assessment of LTp50: An <i>Escherichia coli</i>-Made Chimeric Antigen Targeting S1- and S2-Epitopes from the SARS-CoV-2/BA.5 Spike Protein |
| title_full_unstemmed | Production and Immunogenicity Assessment of LTp50: An <i>Escherichia coli</i>-Made Chimeric Antigen Targeting S1- and S2-Epitopes from the SARS-CoV-2/BA.5 Spike Protein |
| title_short | Production and Immunogenicity Assessment of LTp50: An <i>Escherichia coli</i>-Made Chimeric Antigen Targeting S1- and S2-Epitopes from the SARS-CoV-2/BA.5 Spike Protein |
| title_sort | production and immunogenicity assessment of ltp50 an i escherichia coli i made chimeric antigen targeting s1 and s2 epitopes from the sars cov 2 ba 5 spike protein |
| topic | linear epitopes built-in adjuvant humoral response chimeric antigen COVID-19 |
| url | https://www.mdpi.com/1424-8247/17/3/302 |
| work_keys_str_mv | AT alejandrawongarce productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT omargonzalezortega productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT andrearomeromaldonado productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT arlethmirandalopez productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT marianogarciasoto productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT susanfarfancastro productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT lourdesbetancourtmendiola productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT samapornteeravechyan productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT kanjanasrisutthisamphan productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT mauriciocomasgarcia productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT karlaisolisandrade productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein AT sergiorosalesmendoza productionandimmunogenicityassessmentofltp50aniescherichiacoliimadechimericantigentargetings1ands2epitopesfromthesarscov2ba5spikeprotein |