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J Biotechnol. 2013 May 10;165(1):69-75. doi: 10.1016/j.jbiotec.2013.02.013. Epub 2013 Mar 16.

Display of Neospora caninum surface protein related sequence 2 on Rous sarcoma virus-derived gag protein virus-like particles.

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  • 1Laboratory of Biotechnology, Integrated Bioscience Section, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.

Abstract

Virus-like particles (VLPs) displaying antigen have been increasingly recognized as a potential vaccine in the livestock industry. In this study, Neospora caninum surface protein related sequence (NcSRS)2 was displayed on the surface of Rous sarcoma virus group-antigen protein (RSV-gag) VLPs. Two types of Bombyx mori nucleopolyhedrovirus (BmNPV) bacmids, encoding RSV-gag and NcSRS2 genes, were co-injected into silkworm larvae to produce VLPs-NcSRS2. At 7 days post-injection, VLPs-NcSRS2 were collected from hemolymph and purified. The antigenicity of the purified protein was confirmed by enzyme-linked immunosorbent assay (ELISA) using neosporosis-positive bovine serum. ELISA revealed that ~0.16μg rNcSRS2 was displayed per 1μg VLPs-NcSRS2. To develop an antibody specific for VLPs-NcSRS2, purified VLPs-NcSRS2 were used to immunize mice in a three-dose regimen without adjuvant and the production of antibodies was confirmed in serum samples. By using a silkworm expression system, we demonstrated the display, expression and immunization of neosporosis-targeting membrane proteins, which are vaccine candidates for neosporosis.

Copyright © 2013 Elsevier B.V. All rights reserved.

PMID:
23511652
[PubMed - indexed for MEDLINE]
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