Artículos de revistas
Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system
Fecha
2014-03Registro en:
Targovnik, Alexandra Marisa; Villaverde, Marcela Solange; Arregui, Mariana Bernadett; Fogar, Mariela A.; Taboga, Oscar Alberto; et al.; Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system; Elsevier; Process Biochemistry; 49; 6; 3-2014; 917-926
1359-5113
1873-3298
Autor
Targovnik, Alexandra Marisa
Villaverde, Marcela Solange
Arregui, Mariana Bernadett
Fogar, Mariela A.
Taboga, Oscar Alberto
Glikin, Gerardo Claudio
Finocchiaro, Liliana Maria Elena
Cascone, Osvaldo
Miranda, Maria Victoria
Resumen
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.