Classical swine fever virus E2 glycoprotein antigen produced in adenovirally transduced PK-15 cells confers complete protection in pigs upon viral challenge

Vaccine
Oliberto SanchezJorge R Toledo

Abstract

E2 is the major envelope glycoprotein present on the outer surface of the classical swine fever virus (CSFV). It is exposed as a homodimer originated by disulfide linkage and represents an important target for the induction of neutralizing immune responses against the viral infection. The E2his glycoprotein nucleotide sequence used in this work contains the CSFV E2 extracellular domain preceded by the tissue plasminogen signal peptide and a hexa-histidine tag in the 3' terminus. The recombinant antigen was produced at a range of 120-150 microg/mL in the culture media of epithelial kidney pig cells, transduced with a replication defective adenoviral vector (Ad-E2his) generated by means of cloning the E2his sequence in the vector genome. The glycoprotein was obtained from clarified culture media as a homodimer of 110 kDa with purity over 95% after a single affinity chromatography step in Ni-NTA Agarose column. The E2his characterization by lectin-specific binding assay showed the presence of N-linked oligosaccharides of both hybrid and complex types. The protective capacity of E2his was demonstrated in two immunization and challenge experiments in pigs using doses of 15 or 30 microg of the glycoprotein, emulsified in Freund's adj...Continue Reading

References

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Aug 29, 2006·Vaccine·Xiao-Nan Dong, Ying-Hua Chen

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Citations

Sep 6, 2008·Developmental and Comparative Immunology·Kenneth C McCullough, Artur Summerfield
Sep 17, 2015·Protein Expression and Purification·Camila E Beltrán-OrtizOliberto Sánchez
Jan 11, 2017·Veterinary Microbiology·Sandra BlomeMartin Beer
Mar 11, 2021·Veterinary Immunology and Immunopathology·Marisela Suárez-PedrosoMario Pablo Estrada-García
Dec 20, 2008·Journal of Proteome Research·Raquel MontesinoJose A Cremata
May 13, 2010·Archives of Biochemistry and Biophysics·Raquel MontesinoJosé A Cremata

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