{"id":14284,"date":"2021-03-29T09:35:25","date_gmt":"2021-03-29T09:35:25","guid":{"rendered":"https:\/\/clinlabint.com\/?p=14284"},"modified":"2021-04-02T09:42:02","modified_gmt":"2021-04-02T09:42:02","slug":"separation-and-characterization-of-viruses-using-multi-detector-field-flow-fractionation","status":"publish","type":"post","link":"https:\/\/clinlabint.com\/separation-and-characterization-of-viruses-using-multi-detector-field-flow-fractionation\/","title":{"rendered":"Separation and characterization of viruses using multi-detector field-flow fractionation"},"content":{"rendered":"
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\r\n\"Bio-Rad<\/a>\r\n<\/p>\n<\/div><\/section><\/div>

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Separation and characterization of viruses using multi-detector field-flow fractionation<\/h1>\/ in Featured Articles<\/a> <\/span><\/span><\/header>\n<\/div><\/section>
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by Dr Robert Reed and Dr Soheyl Tadjiki<\/em><\/p>\n

Viruses are increasingly used as gene therapy vectors in modern medicine. In order to minimize unwanted side effects such as immunogenicity, accurate determination of viral aggregation status is important. Field-flow fractionation is emerging as one of the most robust techniques for the separation and size measurement of viruses, their fragments, and their aggregates.<\/strong><\/p>\n

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Introduction<\/h3>\n

The global coronavirus pandemic is the biggest story of 2020, thrusting viruses to the forefront of almost every person\u2019s consciousness around the world. Characterization of viruses and the development of vaccines have become the most important scientific pursuits for many labs.<\/p>\n

Vaccines can be complex and may contain many molecular species that span a large size range, from antibodies, virus fragments such as proteins and nucleic acids, polysaccharide\u2013protein complexes and up to large aggregates that are often over 100 nm in diameter. This means these samples are challenging to separate and characterize by column-based chromatography techniques such as size exclusion chromatography.<\/p>\n

\u2018Asymmetrical flow field-flow fractionation\u2019 (AF4) can help researchers in this field as it is a stationary phase-free separation technique for characterization of polydisperse samples from approximately 1 nm to 1 \u03bcm in diameter [1, 2]. AF4 has already shown merit in the characterization of viruses and virus-like particles [3, 4], as well as antibodies and antigens and their aggregates and agglomerates [5, 6].<\/p>\n

A schematic for the AF4 channel is shown in Figure 1. The combination of cross flow and channel flow causes size separation over the course of the analysis, with smaller particles eluting to connected detectors before larger particles, including aggregates.<\/p>\n

For characterization of the separated particles or molecules, a variety of detectors can be connected to the AF4, including dynamic light scattering and multi-angle light scattering (MALS) for size measurement, with the latter also providing data on molecular weight when combined with a concentration detector such as UV\/Vis or refractive index.<\/p>\n

In this article, AF4 coupled with multiple detectors was used to separate and characterize:<\/p>\n