OBJECTIVETo establish a method for pilot-scale production and quality control of multiepitope hepatitis B virus (HBV) DNA vaccine (PVAX-HS).

METHODSRecombinant DH5alpha/pVAX-HS was obtained by fed-batch fermentation, and the plasmid was extracted by alkaline lysis and concentrated by ultrafiltration. The plasmid DNA was purified by a three-step column chromatography to obtain the DNA vaccine, and quality control tests were performed on the final product.

RESULTSThe quantity of the fed-batch product reached 50-60 g/L, and the final plasmid output was 1.0 mg per gram of the bacteria. The quality of the DNA vaccine met the requirements for medical use.

CONCLUSIONA simple and stable procedure was established for pilot-scale production of multiepitope HBV DNA vaccine, which allows potential large-scale production of the DNA vaccine.