Viruses and virus-like particles have become very attractive formats for vaccines, gene- or oncotherapy. The bioprocessing of these bionanoparticles is challenging due presence of  other particulate impurities, such as microvesicles, exosomes, chromatin and other vesicles. These are very similar in size with a size range between 30 and 300 nm and similar charge and surface properties. Therefore, high-resolution methods are required to separate them. Charged and functionalized non-woven fiber materials possess optimal features to separate nanoparticles from other process related impurities such as host cell proteins and small host cell DNA. The bio-nanoparticle can be loaded at high salt concentration and HCP and small DNA does not bind,  while the particles bind in high capacity.  Here, we present a capture method for recombinant viruses and virus like particles directly from clarified cell culture supernatant based on functionalized non-woven filters with anion-exchange functional groups. The non-wovens exhibit fast loading kinetics without  mass transfer limitation, and have a high capacity and a high virus recovery. The separation of different particle fractions is demonstrated by immune assay, high resolution cryotomography and multiangle light scattering methods. These filters are autoclavable, and they can be integrated into a sterile process and scale up to a size equivalent to several hundred liters.

Virus and virus like particle purification by fiber chromatography

ACS Spring 2022

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