title: Chromatographically-purified capsid proteins of red-spotted grouper nervous necrosis virus expressed in Saccharomyces cerevisiae form virus-like particles.
abstract: 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 25nm, 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.
authors.affiliation: College of Pharmacy, Chung-Ang University, 221 Huksuk-Dong, Dongjak-Gu, Seoul 156-756, South Korea.
authors.forename: Yoo Ri
authors.initials: YR
authors.lastname: Choi
authors.forename: Hyoung Jin
authors.initials: HJ
authors.lastname: Kim
authors.forename: Jae Young
authors.initials: JY
authors.lastname: Lee
authors.forename: Hyun Ah
authors.initials: HA
authors.lastname: Kang
authors.forename: Hong-Jin
authors.initials: HJ
authors.lastname: Kim
mesh_terms: D000818:Animals; D001492:Bass / Q000821:virology*; D002213:Capsid / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D003001:Cloning, Molecular; D005260:Female; D005393:Fish Diseases / Q000276:immunology / Q000517:prevention & control*; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D030381:Nodaviridae / Q000235:genetics* / Q000276:immunology; D012327:RNA Virus Infections / Q000276:immunology / Q000517:prevention & control*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D058425:Vaccines, Virus-Like Particle / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D014771:Virion / Q000235:genetics / Q000378:metabolism
chemical_list: D036022:Capsid Proteins; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D058425:Vaccines, Virus-Like Particle