Enveloped virus- like particles are budded from the host cell into the cell culture supernatant during the production process. The correctly assembled VLPs contain the membrane proteins of the host cell. In addition, other similar sized extracellular particles, such as microvesicles or exosomes are also released from the cell into the culture supernatant. Additionally, the feedstock is contaminated with DNA, free viral or host cell proteins and aberrantly assembled virus-like structures that could contain genetic material. An efficient purification strategy based on connective media as stationary phase to separate VLPs from these main impurities has been developed. The VLPs were directly captured from to the clarified cell culture supernatant and separated into two fractions. As a model, we studied enveloped HIV-1 gag VLPs expressed in CHO cells and influenza VLP expressed in insect cells. They are approximately 100 to 150 nm in size, mimic viral structures and are able to elicit strong immune response. Consequently, they are promising vaccine candidates or can be used in immuno- or gene therapy. The developed process is suited for a platform production of enveloped VLPs.

High resolution separation of virus-like particles and extracellular particles by convective media

253rd National Meeting (2017)

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