Journal
PROTEIN EXPRESSION AND PURIFICATION
Volume 89, Issue 2, Pages 162-168Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2013.03.007
Keywords
Nodavirus; Capsid protein; Virus-like particles (VLPs); Heparin chromatography; Saccharomyces cerevisiae
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Funding
- Korean Ministry of Rural Development Administration (Next-Generation BioGreen 21 Programme) [ADFC2013-PJ009586]
- Rural Development Administration (RDA), Republic of Korea [PJ009586022013] Funding Source: Korea Institute of Science & Technology Information (KISTI), National Science & Technology Information Service (NTIS)
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Nervous necrosis viruses (NNVs) cause mass mortality of marine fish, leading to large economic losses for aquaculturists. A promising vaccine candidate for preventing NNV infection is the NNV virus-like particle (VLP), which is a structure resulting from assembly of recombinant NNV capsid protein. NNV capsid proteins have been expressed in insect cells and the Escherichia coli expression system, and purified by non-scalable protocols such as ultracentrifugation on sucrose and cesium chloride density gradients. In this study, we expressed red-spotted grouper NNV (RGNNV) capsid proteins in Saccharomyces cerevisiae (S. cerevisiae) and developed a chromatography-based method with potential for large-scale vaccine production. The RGNNV capsid protein was successfully purified by a single-step of heparin chromatography. Transmission electron microscopy (TEM) confirmed the high quality of the purified RGNNV capsid protein: it was in the form of VLPs with mean diameters of 25 nm, in homogeneous suspension without any aggregation. Moreover, the RGNNV capsid protein elicited anti-RGNNV capsid protein antibodies in mice. We suggest that RGNNV capsid protein expressed in S. cerevisiae and purified by heparin chromatography, is of sufficient quality for use as a vaccine. (c) 2013 Elsevier Inc. All rights reserved.
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