流感病毒裂解工艺的优化

发布时间：2019-01-29 20:45

【摘要】：目的探讨影响流感病毒裂解的因素,优化流感病毒裂解工艺。方法分别在不同裂解时间(Triton X-100裂解1、2、3和4 h)、不同终浓度的裂解剂(终浓度为0.5%、0.6%、0.7%和0.8%的Triton X-100)、不同总蛋白浓度(2 000、3 000和4 000μg/ml)及不同的离子强度[终浓度为0.01、0.05、0.1和0.5 mol/L的PBS(p H 7.2)]下,对流感病毒进行裂解,电镜下观察病毒裂解效果,同时采用免疫单扩散法检测血凝素含量,Lowry法检测总蛋白含量,计算蛋白含量/血凝素含量比值(裂解后病毒纯度)及血凝素回收率,确定最佳裂解工艺参数。结果当裂解前病毒纯化液蛋白含量范围为2 000~3 000μg/ml,PBS终浓度为0.1 mol/L时,利用终浓度为0.7%~0.8%的Triton X-100裂解流感病毒2 h,可获得最佳裂解效果。结论确定了流感病毒最佳裂解参数,优化了流感病毒裂解疫苗生产中的裂解工艺。
[Abstract]:Objective to study the factors affecting influenza virus cleavage and optimize the process of influenza virus cleavage. Methods at different pyrolysis time (Triton X-100), the final concentration of Triton X-100 was 0. 5% and 0. 8%, respectively, and the final concentration was 0. 5%, 0. 6% and 0. 8% Triton X-100 at different final concentrations (0. 5%, 0. 6% and 0. 8% Triton X-100), respectively. Different concentrations of total protein (2 000 渭 g/ml and 4 000 渭 g/ml) and different ionic intensities (final concentrations of 0. 01% 0. 05% and 0. 5 mol/L of PBS (p H 7.2) were used to break down the influenza virus. The effect of virus cleavage was observed by electron microscope, hemagglutinin content was detected by immunodiffusion method, total protein content was detected by Lowry method, the ratio of protein content to hemagglutinin content (virus purity after lysis) and the recovery rate of hemagglutinin were calculated. Determine the optimum pyrolysis process parameters. Results when the protein content of purified prelytic virus was in the range of 2 000 渭 g / ml PBS for 0. 1 mol/L, the best lytic effect could be obtained by using 0. 7% Triton X-100 with 0.8% final concentration for 2 h. Conclusion the optimal lysis parameters of influenza virus were determined and the lysis process of influenza virus lytic vaccine was optimized.
【作者单位】：长春生物制品研究所有限责任公司疫苗六室;
【基金】：国家科技重大专项(2012ZX09201301-006) 吉林省医药产业发展专项资金(YYZX201112)
【分类号】：R392

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