Production Process Of Hemorrhagic Fever With Renal Syndrome Bivalent Purified Vaccine Improvement

Objective Optimizing the production process conditions of bivalent hemorrhagic fever with renal syndrome vaccine.Methods 1.To investigate the disinfection effect of benzalkonium bromide disinfectant on gerbils before dissection through microbial limit test.2.11-20 day old gerbils were dissected,kidneys were taken,and cells were cultured,and the optimal age of gerbils was confirmed by observing the adhesion and quantity of cells.3.Using the Box-Behnken test,design a 3-factor 3-level response surface test scheme of inoculum MOI,culture temperature and culture days,respectively measure the production parameters of type Ⅰ and type Ⅱ virus harvest liquid,and optimize the process conditions.4.Use ultrafiltration chromatography to sample the first and second peaks of the concentrated liquid chromatography to detect the antigen content,and study whether there are antigens in other peaks during the chromatography5.Antigen detection method was used to measure type Ⅰ and type Ⅱ hemorrhagic fever stock solution after sterilizing and filtering,comparing before and after washing with PBS,and detecting antigen recovery rate.Results 1.Microbial limit test results show that benzalkonium bromide disinfectant can control the microbial level on the surface of gerbils after disinfection to the standard of purified water,meeting the cleanliness requirements of vaccine production.2.The results of cell attachment rate experiments show that the kidney cells of 12-day-old gerbils are the optimal age for the preparation of vaccine production cells.3.The results of response surface experiments showed that the optimal production parameters of type Ⅰ virus harvest liquid were inoculum volume MOI 0.02,virus culture for 5 days,and culture temperature at 35℃.The optimal production parameters of type Ⅱ virus harvest liquid were inoculum MOI 0.02,virus culture for 6 days,and culture temperature at 36℃.The fitting models of virus titer in the harvest fluid of type I and type II hemorrhagic fever were constructed by variance analysis.4.The results of ultrafiltration chromatography experiments show that the protein removal rate of the eluate is above 90%,and the second peak of the concentrated liquid chromatography contains a small amount of antigen.5.The results of the antigen recovery test show that the antigen recovery rate of the type I and type II purified liquid after the original solution is filtered by the filter capsule increases from 20%to more than 80%.Conclusion This article studies and optimizes the production process of bivalent hemorrhagic fever with renal syndrome vaccine,determines the optimal production conditions for type I and type II virus harvesting solution and stock solution,and provides important experimental technical reference for the improvement and application of production technology for cultivating and preparing the vaccine.