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首页> 外文期刊>Physical chemistry chemical physics: PCCP >Impact of the virus purification protocol on aggregation and electrokinetics of MS2 phages and corresponding virus-like particles
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Impact of the virus purification protocol on aggregation and electrokinetics of MS2 phages and corresponding virus-like particles

机译：病毒纯化方案对MS2噬菌体及相应病毒样颗粒聚集和电动的影响

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Previous experimental and theoretical studies have established that electrokinetic and aggregation properties of soft MS2 phages are not only governed by the physico-chemical features of their proteinaceous outer surface but are also significantly impacted by those of their inner RNA component (Dika et al. Appl. Environ. Microbiol., 2011, 14, 4939-4948). These conclusions contradict the recent findings of Nguyen et a/. (Soft Matter, 2011, 7, 10449-10456) who reported identical electrokinetic and aggregation characteristics for MS2 and corresponding virus like particles (VLPs) that lack the internal RNA component. We demonstrate here that this contradiction originates from the different purification methods adopted prior to measurements. More generally, we show that stability and electrohydrodynamics of viruses differ according to purification by (i) dialysis, (ii) isopycnic centrifugation in the cesium chloride gradient, and (iii) precipitation using polyethylene glycol (PEG). Methods (i) and (iii) lead to aggregation of MS2 phages at pH < 4 and pH < 6 in 1-100 mM NaN03 solutions, respectively, while under such conditions aggregation is not observed for MS2 and VLP suspensions prepared according to (ii). In addition, VLPs prepared following methods (i) and (iii) aggregate only at the isoelectric point (pH ～ 3-4) in 1 mM NaNO3 solution. Electrophoretic mobility data of stable MS2 and VLP particles were further examined using a recent formalism for electrokinetics of soft multilayered colloids. The analysis qualitatively shows how the purification protocol may affect either the outer particle surface properties and/or the inner particle content. Finally, the non-DLVO aggregation behavior of MS2 and VLPs purified via the above protocols is discussed in terms of the possible change in corresponding interparticular interactions.
机译：先前的实验和理论研究已经确定，软MS2噬菌体的电动和聚集特性不仅受其蛋白质外表面的物理化学特征支配，而且还受到其内部RNA成分的显着影响（Dika等人，Appl。环境微生物学，2011，14，4939-4948）。这些结论与Nguyen等人的最新发现相矛盾。 （Soft Matter，2011，7，10449-10456）报告了MS2和缺少内部RNA成分的相应病毒样颗粒（VLP）的相同的电动和聚集特征。我们在这里证明了这种矛盾源自测量之前采用的不同纯化方法。更一般而言，我们显示病毒的稳定性和电动动力学因通过（i）透析，（ii）氯化铯梯度中的等密度离心和（iii）使用聚乙二醇（PEG）沉淀纯化而不同。方法（i）和（iii）分别导致1-100 mM NaNO3溶液中pH <4和pH <6的MS2噬菌体聚集，而在这种条件下，根据（ii）制备的MS2和VLP悬浮液未观察到聚集）。此外，按照方法（i）和（iii）制备的VLP仅在1 mM NaNO3溶液的等电点（pH〜3-4）下聚集。稳定的MS2和VLP颗粒的电泳迁移率数据使用最新的形式学方法对软多层胶体的电动学进行了进一步研究。该分析定性地显示了纯化方案如何影响外部颗粒表面性质和/或内部颗粒含量。最后，根据相应微粒间相互作用的可能变化，讨论了通过上述协议纯化的MS2和VLP的非DLVO聚集行为。

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