Production of biologically active feline interferon beta in insect larvae using a recombinant baculovirus
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Feline interferon beta is a cytokine that belongs to the type I IFN family, with antitumor, antiviral and immunomodulatory functions. In this work, recombinant feline interferon beta (rFeIFNβ) was expressed in insect larvae that constitute important agronomic plagues. rFeIFNβ accumulated in the hemolymph of Spodoptera frugiperda larvae infected with recombinant baculovirus and was purified by Blue-Sepharose chromatography directly from larval homogenates on day 4 post-infection. rFeIFNβ was recovered after purification with a specific activity of 1 × 106 IU mg−1. By this method, we obtained 8.9 × 104 IU of purified rFeIFNβ per larva. The product was biologically active in vitro, with an antiviral activity of 9.5 × 104 IU mL−1, as well as a potent antitumor activity comparable to that of the commercial FeIFNω. The glycosylation of rFeIFNβ was confirmed by peptide-N-glycosidase F digestion. Our findings provide a cost-effective platform for large-scale rFeIFNβ production in laboratory research or veterinary medicine applications.
KeywordsFeline interferon beta Baculovirus Insect larvae Veterinary medicine
Autographa californica multiple nucleopolyhedrovirus
Recombinant feline interferon
Fetal bovine serum
Vesicular stomatitis virus
Modified Eagle’s medium
Multiplicity of infection
This work was supported by Grants from Agencia Nacional de Promoción Científica y Tecnológica de Argentina and from Universidad de Buenos Aires (PICT 2014-3350; UBACyT 2016-20020150100145BA). AMT, MSV, FJW and MVM are career researchers of the Consejo Nacional de Investigaciones Científicas y Técnicas de Argentina (CONICET). MBA is a research fellow of CONICET. GJM is a research fellow of Agencia de Promoción Científica y Tecnológica (ANPCyT).
MBA worked out almost all of the technical details and performed the numerical calculations; GJM and IS contributed with analytical methods; MSV performed antitumor activity assays; FJW contributed in downstream processing; AMT contributed in discussion; MVM was involved in planning and supervised the work. All authors discussed the results and commented on the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest in the publication.
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