Yan Guo1, Chang-ye Hui. 1. National Hepatobiliary and Enteric Surgery Research Center of Ministry of Health, Central South University, Changsha, 410078, China. yanguo615@163.com
Abstract
OBJECTIVE: To establish a method for pilot-scale production and quality control of multiepitope hepatitis B virus (HBV) DNA vaccine (PVAX-HS). METHODS: Recombinant 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. RESULTS: The 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. CONCLUSION: A 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.
OBJECTIVE: To establish a method for pilot-scale production and quality control of multiepitope hepatitis B virus (HBV) DNA vaccine (PVAX-HS). METHODS: Recombinant 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. RESULTS: The 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. CONCLUSION: A 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.
Authors: Fuad A A Alssamei; Najla A Al-Sonboli; Fawzi A Alkumaim; Nader S Alsayaad; Mohammed S Al-Ahdal; Tarig B Higazi; Atif A Elagib Journal: Hepat Res Treat Date: 2017-03-06