不同截留分子量超滤膜包浓缩狂犬病病毒的效果

Concentration of rabies virus by ultrafiltration membranes with various molecular weight cutoff values

目的比较不同截留分子量超滤膜包浓缩狂犬病病毒(rabies virus,RV)的效果。方法将RV固定毒3aG-V株按0.01 MOI比例接种至Vero细胞中,制备RV原液,分别用截留分子量100 KD和300 KD的膜包超滤浓缩60倍,β-丙内酯灭活后,验证病毒液浓缩过程膜下液的病毒灭活效果;采用Sepharose 4FF凝胶层析纯化灭活的病毒浓缩液,Lowry法测定病毒浓缩液和纯化液中的蛋白质含量;酶联免疫法测定病毒浓缩过程膜下液、浓缩液和纯化液中的RV抗原含量。结果 100 KD和300 KD的超滤膜包均能有效截流RV,3次试验膜下液RV抗原含量均为0 IU/ml,观察期内小鼠均健存。300 KD膜包超滤的浓缩液和纯化液比100 KD超滤膜包超滤的浓缩液和纯化液中杂蛋白含量低,抗原含量高。结论截留分子量300 KD的膜包比100 KD的膜包更适用于RV的浓缩。

Objective To compare the efficacies of concentration of rabies virus （RV） by uhrafiltration membranes with various molecular weight cutoff values. Methods Fixed RV 3aG-V strain was inoculated into Veto cells in a MOI of 0. 01. The prepared bulk of RV was concentrated by 60 folds by uhrafihration membranes with molecular weight cutoff values of 100 and 300 KD respectively, then inactivated with β propiolactone. The liquid flowing through the membranes was verified for inactivation efficacy of RV during concentration. The concentrated RV bulk after inactivation was purified by Sepharose 4FF gel chromatography. The protein contents in concentrated and purified RV liquids were determined by Lowry method. Tile antigen contents in liquid flowing through the membranes as well as concentrated and purified RV liq- uids were deterulined by ELISA. Results RV was intercepted effectively by the ultrafihration membranes with molecular weight cutoff values of 100 and 300 KD. All the RV antigen contents in liquid flowing through the membranes in three tests were 0 IU / ml. All the mice survived the observation period. However, the foreign protein content was lower, while the antigen content was higher in the concentrated and purified RV liquids by 300 KD ultrafiltration membrane than those by 100 KD uhrafihration membrane. Conclusion As compared 100 KD uhrafihration membrane, 300 KD uhrafiltraion membrane was more suitable for concentration of RV.