Preliminary Study On Cleaning Verification Of The Pilot Plant For Rotavirus Vaccine Production

High-pressure homogenizer was chosen to verify the cleaning methods and procedures as it was used in rotavirus manufacturing process in pilot according risk assessment.At the same time,this study also confirmed the effectiveness of several disinfectants currently in use and soon to be used.The above work accumulates experience and data for the cleaning verification work of large-scale production of hexavalent rotavirus vaccine.Cellular immunofluorescence is a method for measuring intracellular rotavirus which can bind mouse monoclonal antibody(Mab)against rotavirus.The Mab can bind fluorescein-labeled antiserum against mouse IgG which can be irradiated by ultraviolet to be detected.When cells infected with rotavirus,the virus multiplies in cells and can be traced by Cellular immunofluorescence.According to the relevant requirements of Chinese Pharmacopoeia,this method should be validated including specificity,detection limit and durability.The results show that the method is suitable for the detection of rotavirus residue in the process of high-pressure homogenizer cleaning verification.During the implementation of equipment cleaning verification,due to the characteristics of fully enclosed equipment,the final rinse water sampling was selected.Test items include physical appearance,residual active substances(live virus and Total Organic Carbon content),residual detergent(pH value and conductivity),bacterial endotoxins and bioburden of microbials.The test results show that the cleaning agent and cleaning procedure are correct and effective,which can prevent contamination and cross contamination and guarantee the quality of the final product.The chemical disinfectant is suitable for various site.The disinfection efficiency depends on the incubation time and concentration.In this study,the disinfection efficacies of 75% ethanol,70% isopropyl alcohol,1% 84 disinfectant,0.8% basic phenol,0.4% acid phenol,0.1% chlorpheniramine and spore killers from two manufacturers were confirmed.The results showed that except for the spore killing agent produced by Contec Cleanroom Technology Co.,Ltd.the other seven disinfectants all had good killing effect on the bacterial and fungal propagators under the concentration and 3 action time requirements specified in the product specification.The log values of killing microbials in each experiment are >5.The sporicidal agent produced by Contec had a good killing effect on bacterial propagules.It can effectively kill fungal propagules after prolonging the action time.