Improvised two-step elution of chromatographic purification of grouper’s iridovirus plasmid-based vaccine by monolithic adsorbent

Tamar KANSIL, Clarence M. ONGKUDON, Kenneth F. RODRIGUES & Yi Wei CHAN

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Keywords: Grouper’s iridovirus plasmid-based vaccine; two-step elution;  monolithic purification; chromatography

A b s t r a c t
Vaccines have significantly reduced the antibiotics usage especially in the aquaculture industry. Plasmid-based vaccine offers potential effective immunity against the Grouper Iridovirus and Grouper Nervous Necrosis Virus in groupers. Chromatographic purification system is crucial for the efficient recovery of plasmid DNA vaccine construct. With monolith technology, a platform for the vaccine purification has been developed to obtain a high-throughput production of plasmid DNA vaccine in economic manner and time less. The monolith was prepared via free radical co-polymerization of glycidyl methacrylate GMA and ethylene glycol dimethacrylate EDMA in the presence of cyclohexanol as porogen at 70% porogen concentration which gave a minimum heat build-up and homogenous pore size distribution. Monolithic based purification concluded that 0.1 M NaCl as a running buffer coupled with 0.9 M of sulphate salt solution and a flow rate of 1.0 mL/min used for separation are found to be able to produce a reasonably pure DNA. First peak elution is eluting all the RNA leaving DNA alone to be eluted in the second peak. The result of the studies will advance knowledge in the control of disease in the aquaculture industry using plasmid based vaccination and high throughput plasmid purification process.

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