Basic and applied virus research requires specimens that are purified to high homogeneity. Thus, there is much interest in the efficient production and purification of viruses and their subassemblies. Advances in the production steps have shifted the bottle neck of the process to the purification. Nonetheless, the development of purification techniques for different viruses is challenging due to the complex biological nature of the infected cell cultures as well as the biophysical and -chemical differences in the virus particles. We used bacteriophage ϕ6 as a model virus in our attempts to provide a new purification method for enveloped viruses. We compared asymmetrical flow field-flow fractionation (AF4)-based virus purification method to the well-established ultracentrifugation-based purification of ϕ6. In addition, binding of ϕ6 virions to monolithic anion exchange columns was tested to evaluate their applicability in concentrating the AF4 purified specimens. Our results show that AF4 enables one-hour purification of infectious enveloped viruses with specific infectivity of ~1 × 10¹³ PFU/mg of protein and ~65–95% yields. Obtained purity was comparable with that obtained using ultracentrifugation, but the yields from AF4 purification were 2–3-fold higher. Importantly, high quality virus preparations could be obtained directly from crude cell lysates. Furthermore, when used in combination with in-line light scattering detectors, AF4 purification could be coupled to simultaneous quality control of obtained virus specimen.

基础和应用病毒研究要求将标本纯化至高度均一。因此，人们对有效生产和纯化病毒及其子组件非常感兴趣。生产步骤的进步已将过程的瓶颈转移到了纯化上。然而，由于被感染的细胞培养物的复杂生物学特性以及病毒颗粒的生物物理和化学差异，针对不同病毒的纯化技术的发展具有挑战性。我们使用噬菌体ϕ6作为模型病毒，试图为包膜病毒提供一种新的纯化方法。我们将基于不对称流场流分级分离（AF4）的病毒纯化方法与已建立的基于超离心的ϕ6纯化方法进行了比较。此外，测试了ϕ6病毒体与整体式阴离子交换柱的结合，以评估其在浓缩AF4纯化样品中的适用性。我们的结果表明，AF4可以一小时纯化具有感染性的包膜病毒，特异性感染率为〜1×10¹³  PFU / mg蛋白质和〜65–95％的产率。获得的纯度与使用超速离心获得的纯度相当，但是从AF4纯化中获得的产率提高了2-3倍。重要的是，可以从粗细胞裂解物中直接获得高质量的病毒制剂。此外，当与在线光散射检测器结合使用时，AF4纯化可与同时对获得的病毒标本进行质量控制相结合。