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

Abstract

Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky's disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA 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 detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.

• Authors: María Soledad Serena, Christoph Geisler, Germán Ernesto Metz, Santiago Gerardo Corva, Eduardo Carlos Mórtola, Alejandra Larsen, Donald L Jarvis, María Gabriela Echeverría
• Journal: Protein expression and purification (Protein Expr Purif) Vol. 90 Issue 1 Pg. 1-8 (Jul 2013) ISSN: 1096-0279 [Electronic] United States
• PMID: 23631926 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013. Published by Elsevier Inc.

Chemical References

• Antibodies, Viral
• Recombinant Proteins
• Viral Envelope Proteins
• glycoprotein E, Suid herpesvirus 1

Topics

• Animals
• Antibodies, Viral (blood)
• Baculoviridae (genetics)
• Enzyme-Linked Immunosorbent Assay
• Herpesvirus 1, Suid (isolation & purification)
• Male
• Mice
• Mice, Inbred BALB C
• Pseudorabies (diagnosis, virology)
• Recombinant Proteins (genetics, isolation & purification, metabolism)
• Sensitivity and Specificity
• Swine
• Viral Envelope Proteins (genetics, isolation & purification, metabolism)