静脉注射人免疫球蛋白工艺中的病毒灭活方法初探

 目的   研究静脉注射人免疫球蛋白（intravenous immunoglobulin，IVIG）工艺中辛酸处理联合20 nm膜过滤的病毒灭活/去除方法。方法   分别将脂包膜Sindbis病毒和伪狂犬病病毒（pseudorabies virus，PRV）加入pH(4.6±0.1)和pH(5.3±0.1)IVIG中间品（辛酸沉淀后上清液），在(7.47±0.39)、(14.47±0.39)和(22.47±0.39) mmol/L辛酸条件下维持(25±1) ℃处理120 min；将非脂包膜猪细小病毒（porcine parovirus，PPV）加入pH(6.0±0.2)IVIG中间品（层析流穿液），用20 nm膜过滤。检测所有样品处理前后的病毒滴度。 结果   (25±1) ℃处理120 min后，pH(4.6±0.1) IVIG中间品经(7.47±0.39)和(14.47±0.39) mmol/L辛酸钠处理后的残余病毒滴度均≤0.50lg，pH(5.3±0.1) IVIG中间品分别经(14.47±0.39)和(22.47±0.39) mmol/L辛酸处理后的残余病毒滴度均≤0.50lg；20 nm膜过滤可使IVIG中间品的PPV滴度下降≥4.00lg。2种处理方法的结果均符合相关规定的要求。结论   在一定条件下，辛酸处理联合20 nm膜过滤可有效灭活/去除IVIG生产过程的相关病毒。

Preliminary study on a virus inactivation method in production process of intravenous immunoglobulin

 Objective   To study a virus inactivation/removel method by caprylic acid treatment combining nanofilm (20 nm) filter in production process of intravenous immunoglobulin (IVIG).  Methods   IVIG intermediates (supernatant after caprylic acid precipitation) at pH(4.6±0.1) and pH(5.3±0.1) were treated respectively with (7.47±0.39), (14.47±0.39) and (22.47±0.39) mmol/L caprylic acid for 120 min at (25±1) ℃ after lipid-enveloped Sindbis virus and pseudorabies virus (PRV) were added. IVIG intermediates (flowing liquid in chromatography) at pH(6.0±0.2) were treated by nanofilm (20 nm) filter after non-lipid-enveloped porcine parvovirus (PPV) was added. The virus titers of all samples were determined before and after treatment.  Results   After treatment for 120 min at (25±1) ℃, the residual virus titers were ≤0.50lg in IVIG intermediates at pH(4.6±0.1) treated with (7.47±0.39) and (14.47±0.39) mmol/L caprylic acid and IVIG intermediates at pH(5.3±0.1) treated respectively with (14.47±0.39) and (22.47±0.39) mmol/L caprylic acid. The titers of residual PPV were reduced by ≥4.00lg after using nanofilm (20 nm) filter. The results for 2 kinds of treatment methods all met requirements.  Conclusion   Caprylic acid treatment combining nanofilm (20 nm) filter can effectively inactivate/remove related viruses in production process of IVIG under certain condition.