Rapid production of a plasmid DNA encoding a malaria vaccine candidate via amino-functionalized poly(GMA-co-EDMA) monolith

  • Danquah M
  • Liu S
  • Ho J
 et al. 
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Malaria is a global health problem; an effective vaccine is urgently needed. Due to the relative poverty and lack of infrastructure in malaria endemic areas, DNA-based vaccines that are stable at ambient temperatures and easy to formulate have great potential. While attention has been focused mainly on antigen selection, vector design and efficacy assessment, the development of a rapid and commercially viable process to manufacture DNA is generally overlooked. We report here a continuous purification technique employing an optimized stationary adsorbent to allow high-vaccine recovery, low-processing time, and, hence, high-productivity. A 40.0 mL monolithic stationary phase was synthesized and functionalized with amino groups from 2-Chloro-N,N-dieth- ylethylamine hydrochloride for anion-exchange isolation of a plasmid DNA (pDNA) that encodes a malaria vaccine candidate, VR1020-PyMSP4/5. Physical characteriza- tion of the monolithic polymer showed a macroporous material with a modal pore di- ameter of 750 nm. The final vaccine product isolated after 3 min elution was homoge- neous supercoiled plasmid with gDNA, RNA and protein levels in keeping with clinical regulatory standards. Toxicological studies of the pVR1020-PyMSP4/5 showed a mini- mum endotoxin level of 0.28 EU/mg pDNA. This cost-effective technique is cGMP com- patible and highly scalable for the production of DNA-based vaccines in commercial quantities, when such vaccines prove to be effective against malaria.

Author-supplied keywords

  • Liquid chromatography
  • Malaria vaccine
  • Methacrylate monolith
  • Plasmid DNA

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  • Michael K. Danquah

  • Shan Liu

  • Jenny Ho

  • Gareth M. Forde

  • Lina Wang

  • Ross L. Coppel

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