Expression and purification of recombinant feline interferon in the baculovirus-insect larvae system

TARGOVNIK, ALEXANDRA M.; VILLAVERDE, MARCELA S.; ARREGUI, MARIANA B.; FOGAR, MARIELA N.; TABOGA, OSCAR; GLIKIN, GERARDO; FINOCCHIARO, LILIANA M. E.; CASCONE, OSVALDO; MIRANDA, MARÍA V.

PROCESS BIOCHEMISTRY - (Print)

ELSEVIER SCI LTD

Lugar: Amsterdam; Año: 2014 vol. 49 p. 917 - 926

1359-5113

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 dayspost-infection (dpi), the concentrations of FeIFN-α7 and FeIFN-α7xArg in suspension culture were (1.28±0.15) x106 UI ml-1 and (1.3±0.2) x106 UI ml-1 respectively. The maximum expression levels of FeIFN-α7 and FeIFN-α7xArg were (3.7±0.2) x106 U ml-1 and (3.5±0.4) x106 U ml-1 at 2 dpi in R. nu larvae and (1.1±0.2) x 106 U ml-1 and (1.0±0.15) x106 U ml-1 at 6 dpi in S. 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.