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

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• 作者：Vipin Kumar Deo^a ; Katsuhiko Yoshimatsu^b ; Takahiro Otsuki^b ; Jinhua Dong^a ; Tatsuya Kato^b ; Enoch Y. Park^a ; ^b ; ^{acypark@ipc.shizuoka.ac.jp}
• 关键词：Display technology ; Neospora caninum surface protein related sequence 2 ; Neosporosis ; Virus-like particle ; Silkworm expression system
• 刊名：Journal of Biotechnology
• 出版年：2013
• 出版时间：10 May, 2013
• 年：2013
• 卷：165
• 期：1
• 页码：69-75
• 全文大小：701 K

文摘

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 ¡«0.16 ¦Ìg rNcSRS2 was displayed per 1 ¦Ìg 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.