Recovery in aqueous two-phase systems of nanoparticulates applied as surrogate mimics for viral gene therapy vectors

A descriptive model and methods for up-scaled process routes for interfacial partition of bioparticles in aqueous two-phase systems

“…It is concluded that the capability of polymer-salt ATPS to distribute bioparticles between the two phases increases with decreasing PEG molecular weight. This has been confirmed for studies with pDNA [25,34] and it appears to be true for other bioparticles such as yeast virus-like particles [35] and adenoviruses [36].…”

Aqueous two-phase recovery of bio-nanoparticles: A miniaturization study for the recovery of bacteriophage T4

“…The non-conventional recovery method, namely ATPS described by Albertson in the mid-1950s [1] has been exploited for the recovery and purification of different bioproducts [1,2]. These include proteins [3], viruses and intact cells [4] virus-like particles [5], inclusion bodies [6], plasmid DNA [7,8] surrogate mimics for viral vectors and adenoviral vectors [9] and even inorganic compounds produced by microorganisms [10]. By speculative comparisons, ATPS might possess a superior volumetric capacity (occupancy per volume of phase) for such nanoparticulate products than adsorptive chromatography density and gradient centrifugation [6,11].…”

“…In response of the limiting amounts of viral vectors for gene therapy applications, washed inclusion bodies as first generation mimics of nanoparticles have been used to predict the partition behavior of adenoviral vectors [9,24]. However, a biological system with similar characteristics to the viral vectors is required.…”

Recent advances in biomolecular process intensification

“…A significant increase in the efficiency of cellular disruption following the addition of detergent and chaotrope has been reported (Bailey et al, 1995), but the addition of detergent complicates subsequent processing and does not eliminate the mechanical disruption step. The use of aqueous two-phase systems for direct recovery of IBs following mechanical disruption has been introduced (Braas et al, 2000), although IBs accumulated at the interface thus complicating their recovery and further processing.…”