Efficient production of porcine circovirus virus-like particles using the nonconventional yeast Kluyveromyces marxianus
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Porcine circovirus type 2 (PCV2) is a ubiquitous virus with high pathogenicity closely associated with the postweaning multisystemic wasting syndrome (PMWS) and porcine circovirus diseases (PCVDs), which caused significant economic losses in the swine industry worldwide every year. The PCV2 virus-like particles (VLPs) are a powerful subunit vaccine that can elicit high immune response due to its native PCV2 virus morphology. The baculovirus expression system is the widely used platform for producing commercial PCV2 VLP vaccines, but its yield and cost limited the development of low-cost vaccines for veterinary applications. Here, we applied a nonconventional yeast Kluyveromyces marxianus to enhance the production of PCV2 VLPs. After codon optimization, the PCV2 Cap protein was expressed in K. marxianus and assemble spontaneously into VLPs. Using a chemically defined medium, we achieved approximately 1.91 g/L of PCV2 VLP antigen in a 5-L bioreactor after high cell density fermentation for 72 h. That yield greatly exceeded to recently reported PCV2 VLPs obtained by baculovirus-insect cell, Escherichia coli and Pichia pastoris. By the means of two-step chromatography, 652.8 mg of PCV2 VLP antigen was obtained from 1 L of the recombinant K. marxianus cell culture. The PCV2 VLPs induced high level of anti-PCV2 IgG antibody in mice serums and decreased the virus titers in both livers and spleens of the challenged mice. These results illustrated that K. marxianus is a powerful yeast for cost-effective production of PCV2 VLP vaccines.
KeywordsPorcine circovirus Capsid protein Virus-like particles Kluyveromyces marxianus
Compliance with ethical standards
This study was supported by the projects of National Natural Science Foundation of China (31770094 and 91731310), National High Technology Research and Development Program of China (2014AA021301), Science and Technology Research Program of Shanghai (18391901800) and Open Research Funds of the State Key Laboratory of Genetic Engineering, Fudan University.
Conflict of interest
The authors declare that they have no competing interests.
All the mice experimental procedures were approved by the Animal Experiment Committee of Fudan University. All applicable international, national, and institutional guidelines for the care and use of animals were strictly followed.
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