Virus-like particles (VLPs) are widely used in medicine, but can be difficult to characterize and isolate from aggregates. In this research, primarily cyclical electrical field-flow fractionation (CyElFFF) coupled with multi-angle light scattering (MALS), and dynamic light scattering (DLS) detectors, was used for the first time to perform size and electrical characterization of three different types of Q beta bacteriophage virus-like particles (VLPs): a blank Q beta bacteriophage which is denoted as VLP and two conjugated ones with different peptides. The CyElFFF results were verified with transmission electron microscopy (TEM). Asymmetrical flow field-flow fractionation (AF4) coupled with MALS was also applied using conditions similar to those used in the CyElFFF experiments, and the results of the two techniques were compared to each other. Using these techniques, the size and electrophoretic characteristics of the fractionated VLPs in CyElFFF were obtained. The results indicate that CyElFFF can be used to obtain a clear distribution of electrophoretic mobilities for each type of VLP. Accordingly, CyElFFF was able to differentially retain and isolate VLPs with high surface electric charge/electrophoretic mobility from the ones with low electric charge/electrophoretic mobility. Regarding the size characterization, the size distribution of the eluted VLPs was obtained using both techniques. CyElFFF was able to identify subpopulations that did not appear in the AF4 results by generating a shoulder peak, whereas AF4 produced a single peak. Different size characteristics of the VLPs appearing in the shoulder peak and the main peak indicate that CyElFFF was able to isolate aggregated VLPs from the monomers partially. Graphical abstract.

中文翻译：

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Q噬菌体病毒样颗粒的表征和差异保留使用循环电场流分馏和不对称流场流分馏。

病毒样颗粒（VLP）在医学中被广泛使用，但可能难以表征和从聚集物中分离出来。在这项研究中，首次使用循环电场流分级分离（CyElFFF），多角度光散射（MALS）和动态光散射（DLS）检测器来进行三种不同类型的尺寸和电表征Qβ噬菌体病毒样颗粒（VLP）的特征：空白的Qβ噬菌体，称为VLP和两个具有不同肽的结合噬菌体。CyElFFF结果通过透射电子显微镜（TEM）验证。还使用类似于CyElFFF实验中所使用的条件，应用了与MALS偶联的非对称流场流动分离（AF4），并将这两种技术的结果进行了比较。使用这些技术，获得了CyElFFF中分离的VLP的大小和电泳特性。结果表明，CyElFFF可用于获得每种VLP类型的电泳迁移率的清晰分布。因此，CyE1FFF能够将具有高表面电荷/电泳迁移率的VLP与具有低表面电荷/电泳迁移率的VLP区别地保留和分离。关于尺寸表征，使用两种技术均获得了洗脱的VLP的尺寸分布。CyElFFF能够通过产生一个肩峰来鉴定未出现在AF4结果中的亚群，而AF4则产生一个峰。出现在肩峰和主峰中的VLP的大小特征不同，表明CyElFFF能够部分地从单体中分离出聚集的VLP。图形概要。