Flow-dependent entrapment of large bioparticles in porous process media

The need for purification of biomolecules extends to larger bioparticles as well. For example, virus purification is required for production of many vaccines and gene delivery vectors, and understanding virus removal in porous media is also important in downstream processing of therapeutic proteins and in purification of water in soils. A convective entrapment mechanism for retention of large bioparticles is discussed here based on retention of such bioparticles in pore constrictions at high enough flow rates, even under non-binding conditions. A simple equation to predict whether such entrapment is expected to occur in a given system is derived based on a Péclet number that is proportional to the flow rate and to the cube of the bioparticle diameter. To test the theory, adenovirus was spiked onto chromatographic beds. As expected from the theory, under non-interacting conditions a progressively larger amount of virus becomes trapped with increasing flow rate. The entrapment is reversible upon flow rate reduction, which, within the proposed model, is based on the possibility of diffusive escape from pore constrictions. This mechanism can be exploited for virus purification or removal, and the theory is also consistent with the anecdotal evidence that monoliths and membranes are more difficult to clean than conventional chromatographic beds, especially at high flow rates.