| Vero cells are popularly applied in the vaccines production. The residual Vero cell in vaccine has potential tumorigenic after inoculation. The risk rate is related with residual amount and size of DNA fragments. Based on this theory, it is very important for improvement quality of vaccine to remove residual DNA.Currently, Vero cells are usually used as media for growth Rabies virus. The harvest will be purified by ultrafiltration and gel filtration chromatography. Even all purified processes completed, the residual DNA amount doesn’t reach a safety level. We tried to adjust parameter of concentration ration, column efficiency, sample volume, salt composition and pH of elution solution, elution liquid flow condition, and so on, to reduce the residual DNA of Vero.Firstly, we optimize the culture condition for cells during virus growth. To reduce the release of Vero cells’DNA, we keep cells in good situation to avoid cells lose and broke while improve the virus harvest titer. Secondly, used suitable ultrafiltration membranes to remove residual DNA and bovine serum albumin without lose antigen. This method could make followed purified steps easier. Thirdly, to meet the standards of Chinese Pharmacopeia (2010vision), we optimize the anion exchange column process, via using appropriate media, to control the antigen lose. The data shows the two previous methods i.e. optimization of culture conditions and utilization300KD ultrafiltration membranes, have good effectiveness, while the third one reducing residual DNA combined with antigen lose. So if want to adopt the last one, the further research should be conducted. |
