Production of Pseudorabies Virus by Vero cells grown on dextran microcarriers

• 1、Jiangnan University school of Biotechnology, Wuxi 214122

Abstract：Microcarriers have been widely applied in the animal vaccine industry because of its advantages, such as high specific surface area and easy to operate, and the market demand is large. At present, commercial microcarriers are mainly imported microcarriers. Due to their high price, it is urgent to independently develop domestic microcarriers. Porcine pseudorabies virus seriously harms the swinery health, causing huge economic losses to farmers. Vaccine immunization prevention is currently the main effective method to control the outbreak of the disease. In this study, self-made microcarriers suspension culture Vero cells were used to produce Porcine Pseudorabies virus, and a safe and reliable virus production process was established. Firstly, the concentration of microcarriers and Vero cell seeding density was optimized in a 125 mL magnetic stirring bottle to create optimal conditions for the self-made microcarriers to cultivate the Vero cells. Secondly, the process parameters of PRV virus culture were optimized, including the time of infection(TOI), multiplicity of infection (MOI), and harvest time. Thirdly, the optimized process parameters were scaled up in a 3 Liter Bioreactor. Based on 72 h cultivation of cells, the highest virus titer LgTCID50omL-1 value of culturing the PRV virus was 7.3 after being infected 72 h at MOI=0.01, which was 6.3 times higher than the reported literature. The economical and practical microcarriers developed in this study will lay the foundation for industrial production of animal vaccines and large-scale production of PRV.

Keywords： Cell engineeringy Vero cell microcarrier Pseudorabies virus 3 L Bioreactor