Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system
Feline interferons (FeIFNs) are cytokines with antiviral, antitumor and immunomodulatory functions used as therapeutic agents in a variety of veterinary diseases. In this work, FeIFN-α7 and FeIFN-α7xArg containing eight residues of arginine were expressed in Sf9 cells and insect larvae. At 4 days po...
| Main Authors: | , , , , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
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2018
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| Online Access: | https://www.sciencedirect.com/science/article/pii/S1359511314001779 http://hdl.handle.net/20.500.12123/2899 https://doi.org/10.1016/j.procbio.2014.03.013 |
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| author | Targovnik, Alexandra Marisa Villaverde, Marcela Solange Arregui, Mariana Bernadett Fogar, Mariela Noemi Taboga, Oscar Alberto Glikin, Gerardo Claudio Finocchiaro, Liliana Maria Elena Cascone, Osvaldo Miranda, Maria Victoria |
| author_browse | Arregui, Mariana Bernadett Cascone, Osvaldo Finocchiaro, Liliana Maria Elena Fogar, Mariela Noemi Glikin, Gerardo Claudio Miranda, Maria Victoria Taboga, Oscar Alberto Targovnik, Alexandra Marisa Villaverde, Marcela Solange |
| author_facet | Targovnik, Alexandra Marisa Villaverde, Marcela Solange Arregui, Mariana Bernadett Fogar, Mariela Noemi Taboga, Oscar Alberto Glikin, Gerardo Claudio Finocchiaro, Liliana Maria Elena Cascone, Osvaldo Miranda, Maria Victoria |
| author_sort | Targovnik, Alexandra Marisa |
| collection | INTA Digital |
| description | Feline interferons (FeIFNs) are cytokines with antiviral, antitumor and immunomodulatory functions used as therapeutic agents in a variety of veterinary diseases. In this work, FeIFN-α7 and FeIFN-α7xArg containing eight residues of arginine were expressed in Sf9 cells and insect larvae. At 4 days post-infection (dpi), the concentrations of FeIFN-α7 and FeIFN-α7xArg in suspension culture were (1.28 ± 0.15) × 106 U ml−1 and (1.3 ± 0.2) × 106 U ml−1 respectively. The maximum expression levels of FeIFN-α7 and FeIFN-α7xArg were (3.7 ± 0.2) × 106 U ml−1 and (3.5 ± 0.4) × 106 U ml−1 at 2 dpi in Rachiplusia nu larvae and (1.1 ± 0.2) × 106 U ml−1 and (1.0 ± 0.15) × 106 U ml−1 at 5 dpi in Spodoptera frugiperda larvae respectively. R. nu was a better host for FeIFN-α7 and FeIFN-α7xArg expression. The 8xArg tag did not affect the biological activity of FeIFN-α7 and was useful to promote the FeIFN-α7xArg adsorption on ion exchange chromatography (IEC), allowing its purification in a single step from supernatant culture and R. nu larvae. FeIFN-α7xArg was purified from the larval extract with a yield of 70% and a purification factor of 25 free of viruses. We conclude that R. nu larvae are new low-cost hosts for the expression of recombinant FeIFN-α7. |
| format | Artículo |
| id | INTA2899 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2018 |
| publishDateRange | 2018 |
| publishDateSort | 2018 |
| record_format | dspace |
| spelling | INTA28992018-07-27T13:29:09Z Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system Targovnik, Alexandra Marisa Villaverde, Marcela Solange Arregui, Mariana Bernadett Fogar, Mariela Noemi Taboga, Oscar Alberto Glikin, Gerardo Claudio Finocchiaro, Liliana Maria Elena Cascone, Osvaldo Miranda, Maria Victoria Interferonas Felidae Gato Citoquinas Baculovirus Cromatografía de Cambio Iónico Purificación Interferons Cats Cytokines Ion Exchange Chromatography Purification Feline interferons (FeIFNs) are cytokines with antiviral, antitumor and immunomodulatory functions used as therapeutic agents in a variety of veterinary diseases. In this work, FeIFN-α7 and FeIFN-α7xArg containing eight residues of arginine were expressed in Sf9 cells and insect larvae. At 4 days post-infection (dpi), the concentrations of FeIFN-α7 and FeIFN-α7xArg in suspension culture were (1.28 ± 0.15) × 106 U ml−1 and (1.3 ± 0.2) × 106 U ml−1 respectively. The maximum expression levels of FeIFN-α7 and FeIFN-α7xArg were (3.7 ± 0.2) × 106 U ml−1 and (3.5 ± 0.4) × 106 U ml−1 at 2 dpi in Rachiplusia nu larvae and (1.1 ± 0.2) × 106 U ml−1 and (1.0 ± 0.15) × 106 U ml−1 at 5 dpi in Spodoptera frugiperda larvae respectively. R. nu was a better host for FeIFN-α7 and FeIFN-α7xArg expression. The 8xArg tag did not affect the biological activity of FeIFN-α7 and was useful to promote the FeIFN-α7xArg adsorption on ion exchange chromatography (IEC), allowing its purification in a single step from supernatant culture and R. nu larvae. FeIFN-α7xArg was purified from the larval extract with a yield of 70% and a purification factor of 25 free of viruses. We conclude that R. nu larvae are new low-cost hosts for the expression of recombinant FeIFN-α7. EEA Sáenz Peña Fil: Targovnik, Alexandra Marisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina Fil: Villaverde, Marcela Solange. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo". Unidad de Transferencia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Arregui, Mariana Bernadett. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina Fil: Fogar, Mariela Noemi. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Sáenz Peña; Argentina Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina Fil: Glikin, Gerardo Claudio. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo". Unidad de Transferencia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Finocchiaro, Liliana Maria Elena. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Oncología "Ángel H. Roffo". Unidad de Transferencia Genética; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cascone, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina Fil: Miranda, Maria Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina 2018-07-27T13:27:01Z 2018-07-27T13:27:01Z 2014-06 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion https://www.sciencedirect.com/science/article/pii/S1359511314001779 http://hdl.handle.net/20.500.12123/2899 1359-5113 https://doi.org/10.1016/j.procbio.2014.03.013 eng info:eu-repo/semantics/restrictedAccess application/pdf Process Biochemistry 49 (6) : 917-926 (June 2014) |
| spellingShingle | Interferonas Felidae Gato Citoquinas Baculovirus Cromatografía de Cambio Iónico Purificación Interferons Cats Cytokines Ion Exchange Chromatography Purification Targovnik, Alexandra Marisa Villaverde, Marcela Solange Arregui, Mariana Bernadett Fogar, Mariela Noemi Taboga, Oscar Alberto Glikin, Gerardo Claudio Finocchiaro, Liliana Maria Elena Cascone, Osvaldo Miranda, Maria Victoria Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system |
| title | Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system |
| title_full | Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system |
| title_fullStr | Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system |
| title_full_unstemmed | Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system |
| title_short | Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system |
| title_sort | expression and purification of recombinant feline interferon in the baculovirus insect larvae system |
| topic | Interferonas Felidae Gato Citoquinas Baculovirus Cromatografía de Cambio Iónico Purificación Interferons Cats Cytokines Ion Exchange Chromatography Purification |
| url | https://www.sciencedirect.com/science/article/pii/S1359511314001779 http://hdl.handle.net/20.500.12123/2899 https://doi.org/10.1016/j.procbio.2014.03.013 |
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