应用纳米膜(DV20_nm)过滤法去除人免疫球蛋白中指示病毒PPV的效果验证

目的: 研究验证纳米膜(DV20 nm)过滤法去除人免疫球蛋白中猪细小病毒(porcine parvovirus,PPV)的去除效果。 方法: 在人免疫球蛋白中加入滴度为6.94 LgTCID₅₀/0.1 mL的PPV为指示病毒,用纳米膜(DV20 nm)进行加压除病毒过滤。根据不同过滤时间点和过滤量进行取样检测样品中残余病毒滴度以评价去除病毒效果。 结果: 当过滤量为5 L·m^-2时,可去除PPV病毒滴度为4.19~4.88 LgTCID₅₀/0.1 mL。过滤量为15 L·m^-2时,去除PPV病毒滴度为3.00~4.19 LgTCID₅₀/0.1 mL。过滤量达到25 L·m^-2时,去除PPV病毒滴度为3.00~3.56 LgTCID₅₀/0.1 mL。 结论: 当总过滤量为60 L·m^-2时,其过滤初期(过滤量5 L·m^-2/时,仅为总量的1/12)去除PPV能力大于4.00 LgTCID₅₀/0.1 mL,而随着过滤量逐渐增加,去除PPV能力逐渐下降。因此,纳米膜过滤法去除病毒必须充分考虑过滤样品的体积、加入指示病毒颗粒大小、产品中蛋白浓度、分子大小、纯度等综合因素,以其达到最佳去除效果。

Objective: To evaluate the effect of nano film filter (DV20 nm) on removing an indicator virus (porcine parvovirus,PPV) in human immunoglobulin. Methods: Human immunoglobulin was added with a titer of 6.94 LgTCID₅₀/0.1 mL PPV as an indicator virus and filtrated with a nano film filer under pressure.Take samples according to the filtered volume at different time points and titrate residual indicator virus to evaluate the removal effect. Results: When an volume of 5 L·m^-2,15 L·m^-2 and 25 L·m^-2 was filtrated,virus could be removed at a titer of 4.19-4.88 LgTCID₅₀/0.1 mL,3.00-4.19 LgTCID₅₀/0.1 mL and 3.00-3.56 LgTCID₅₀/0.1 mL respectively. Conclusion: An early filtration volume (5 L·m^-2,1/12 of total 60 filtration volume ) has a virus removal capacity of more than 4.00 LgTCID₅₀/0.1 mL,however,virus removal capacity is inversely proportional to the filtration volume.Several elements should be incorporated in the Nano film filter testing to optimize the virus removal efficiency,such as virus size,protein size,concentration and purity,etc.

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