Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky?s disease in infected pigs

SERENA M. S; GEISLER CHRISTOPH; METZ G; CORVA S; MORTOLA EC; LARSEN A; JARVIS DONALD L. ; ECHEVERRIA M. G

PROTEIN EXPRESSION AND PURIFICATION

ACADEMIC PRESS INC ELSEVIER SCIENCE

Lugar: Amsterdam; Año: 2013 p. 1 - 8

1046-5928

Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky?s disease (AD), which affects swine herdsworldwide and causes substantial economic losses due to animal mortality and lost productivity. In orderto eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) areongoing in several countries. These eradication programs have generated a currently unmet demand foraffordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinatedpigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologicallyauthentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts toclone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCRenhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. Thecloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reactedstrongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirectELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus,recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detectionof SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinantform of gE as a SHV-1 subunit vaccine