Higuchi Shinsuke, Satou Tatsuki, Uchida Yuki
Japan CMC, Vaccine Operations CMC, Global Vaccine Business Unit, Takeda Pharmaceutical Company Limited, 4720 Takeda, Mitsui, Hikari, Yamaguchi 743-8502, Japan.
MethodsX. 2023 Mar 11;10:102126. doi: 10.1016/j.mex.2023.102126. eCollection 2023.
A virus concentration method is required for viral vaccine manufacturing and virus-related research. However, concentration methods, such as ultracentrifugation, often require capital investment. We report a simple and easy-to-use handheld syringe method for virus concentration using a hollow fiber (HF) filter module, which can be applicable to viruses of different sizes, without incorporating any special machines or reagents. This virus concentration method does not use pumps, which might cause shear stress for virus particles; therefore, it is useful for stress-sensitive virus particles, and virus-like particles, as well as other proteins. The clarified harvest of flavivirus (Zika virus) was concentrated using an HF filter module and compared with a centrifugal ultrafiltration device (CUD) for demonstration of the HF filter method. The HF filter method achieved concentration of the virus solution in less time than the CUD. The yield comparison of the recovered virus solution indicated that recovery from the developed method was comparable to using the CUD, and infectivity was maintained throughout.•The Zika virus was concentrated from 200 mL to 5 mL within 45 min using the HF filter and handheld syringe module method.•The handheld HF filter method may be applicable to stress-sensitive viruses and proteins of different sizes.•The virus concentration process should be conducted in a safety cabinet, which is preferred for virus containment.
病毒疫苗生产和病毒相关研究需要一种病毒浓缩方法。然而,诸如超速离心等浓缩方法通常需要资本投资。我们报告了一种使用中空纤维(HF)过滤模块的简单易用的手持注射器病毒浓缩方法,该方法可适用于不同大小的病毒,无需使用任何特殊机器或试剂。这种病毒浓缩方法不使用可能会对病毒颗粒造成剪切应力的泵;因此,它对压力敏感的病毒颗粒、病毒样颗粒以及其他蛋白质很有用。使用HF过滤模块对黄病毒(寨卡病毒)的澄清收获物进行浓缩,并与离心超滤装置(CUD)进行比较,以展示HF过滤方法。HF过滤方法比CUD在更短的时间内实现了病毒溶液的浓缩。回收病毒溶液的产量比较表明,所开发方法的回收率与使用CUD相当,并且始终保持感染性。
• 使用HF过滤器和手持注射器模块方法,寨卡病毒在45分钟内从200毫升浓缩至5毫升。
• 手持HF过滤方法可能适用于压力敏感病毒和不同大小的蛋白质。
• 病毒浓缩过程应在安全柜中进行,这对于病毒 containment 是首选。