An intermediate-purity virus preparation is achieved by adding a high speed ultracentrifugation step. For example, cellular proteases could degrade virus, while cytokines could alter cell signalling and affect susceptibility to viruses. While this approach is quick and permits simultaneous high-throughput comparisons of a large sundry of viruses, it suffers from high contamination with non-virus factors that may affect experimental outcomes ( Table 1). To generate lysates, cells are commonly lysed by freeze-thaw, debris is removed by low speed centrifugation, and supernatants are then used for experimentation. The most rapid approach involves using crude cell lysates or cell culture media from virus-infected cells. Most laboratories use one of three methods to prepare virus for experimentation.

The in-slurry purification approach offered substantially increased virus purity over crude cell lysates, media, or high-spin preparations and would be especially useful for high-throughput virus screening applications where density gradient ultracentrifugation is not feasible. Capto Core 700 resin was then effectively adapted to a rapid in-slurry pull-out approach for high-throughput purification of reovirus and adenovirus. Core 700 chromatography produced virion purity and infectivity indistinguishable from CsCl density gradient ultracentrifugation as determined by electron microscopy, gel electrophoresis analysis and plaque titration. To overcome this shortcoming, we evaluated a commercially available resin (Capto Core 700) that captures molecules smaller than 700 kDa. Our research pace was limited by the lack of high-throughput virus purification methods that efficiently remove confounding cellular contaminants such as cytokines and proteases. Our laboratory explores mutations in oncolytic reovirus that could improve oncolytic activity, and makes routine use of numerous virus variants, genome reassortants, and reverse engineered mutants. Existing methods to purify viruses such as gradient ultracentrifugation or chromatography have limitations, for example demand for technical expertise or specialized equipment, high time consumption, and restricted capacity.

Viruses are extensively studied as pathogens and exploited as molecular tools and therapeutic agents.

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