Recovery and Manipulation of Nanoparticulate Bioproducts: Relevance to the Up-Scaled Manufacture of Gene Therapy Vectors

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

“…However, the chemical composition of top and bottom phase can vary dramatically, being usually enriched in one of the chemical components [9,24]. To determine the effect of the ATPS chemical components upon the infectivity of the bacteriophage T4, the maximum possible concentrations of the PEG 8000-phosphate and PEG 600-sulphate ATPS components present in the system were tested separately on the bacteriophage T4 infectivity.…”

“…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.…”

Purification of a functional gene therapy vector derived from Moloney murine leukaemia virus using membrane filtration and ceramic hydroxyapatite chromatography

“…However, being based on size, they would ideally be complemented by chromatographic processes that exploit other physicochemical properties (Braas et al, 1996). Consideration of the pore dimensions and particle sizes of common protein purification adsorbents suggests that the adsorption of retroviruses is restricted to the external particle surface area, since the access of retroviruses into pores of diameter 20-200 nm is physically restricted or infeasible (Braas et al, 2000;Lyddiatt et al, l998). By contrast, protein contaminants can access adsorbent surfaces within pores, which present a large surface area for their nonspecific adsorption.…”

Interfacial partition of plasmid DNA in aqueous two-phase systems

“…Inclusion bodies have been successfully separated from cell debris in PEG-salt systems [2]. Viral gene therapy vectors such as retroviruses and adenoviruses and artificial protein particles, produced as surrogate particles for potential vaccine applications, have been separated in PEG-salt systems [3,4]. The recovery of pDNA has been described recently [5,6].…”