Sorption processes in ion-exchange chromatography of viruses

Trilisky, Egor; Lenhoff, Abraham M.

doi:10.1016/j.chroma.2006.12.094

Cited by 78 publications

(80 citation statements)

References 45 publications

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“…(2). Trilisky et al [14] proposed that large biomolecules adsorb virtually irreversible due to their size and therefore the assumption of equilibrium is no longer given. They introduce a model where the rate of adsorption is described in terms of the fractional approach to capacity…”

Section: Batch Uptake Rate Described In Terms Of the Fractional Appromentioning

confidence: 99%

“…Several speculations concerning adsorption of DNA on AEC may complicate a precise mathematical prediction. Due to the very high charge density of the phosphate backbone the binding may be considered as virtually irreversible and solid diffusion does not occur [14]. Intra-as well as intermolecular electrostatic interactions may influence the adsorption mechanism tremendously in a yet not well-understood manner.…”

Section: Introductionmentioning

confidence: 99%

“…The adsorption process can basically be characterized by uptake kinetics. Several attempts had already been made to model the uptake behaviour of large biomolecules such as pDNA and viruses on AEC media [14,15]. Models derived from protein chromatography only partially provide a good approximation.…”

Section: Introductionmentioning

confidence: 99%

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Adsorption of plasmid DNA on anion exchange chromatography media

Tarmann

Jungbauer²

2008

J of Separation Science

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Original PaperAdsorption of plasmid DNA on anion exchange chromatography media Anion exchange chromatography (AEC) is a useful and effective tool for DNA purification, but due to average pore sizes between 40 and 100 nm most AEC resins lack truly useful binding capacities for plasmid DNA (pDNA). Equilibrium binding capacities and uptake kinetics of AEC media including conventional media (Source 30 Q, Q Sepharose HP), a polymer grafted medium (Fractogel EMD DEAE (M)), media with large pores (Celbeads DEAE, PL SAX 4000 30 lm) and a monolithic medium (CIM-DEAE) were investigated by batch uptake or shallow bed experiments at two salt concentrations. Theoretical and experimental binding capacities suggest that the shape of the pDNA molecule can be described by a rod with a length to diameter ratio of 20:1 and that the molecule binds in upright position. The arrangement of DNA like a brush at the surface can be considered as entropy driven, kind of selfassembly process which is inherent to highly and uniformly charged DNA molecules. The initial phase of adsorption is very fast and levels off, associated with a change in mass transfer mechanism. Feed concentrations higher than 0.1 mg/mL pDNA pronounce this effect. Monolithic media showed the fastest adsorption rate and highest binding capacity with 13 mg pDNA per mL.

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Section: Batch Uptake Rate Described In Terms Of the Fractional Appromentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Adsorption of plasmid DNA on anion exchange chromatography media

Tarmann

Jungbauer²

2008

J of Separation Science

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“…In order to support these exploratory experiments, similar tests were performed with another important viral vector in clinical applications: human Ad5, a non-enveloped virus with an equivalent particle diameter of approximately 90-100 nm (Trilisky and Lenhoff, 2007). Even with a smaller overall size compared to rBV, the maximum adsorption of these particles is not affected by the decrease in ligand density (Fig.…”

Section: Impact Of Ligand Density On the Adsorption Capacity Of Deae-mentioning

confidence: 98%

Impact of ligand density on the optimization of ion‐exchange membrane chromatography for viral vector purification

Vicente

Faber

Alves

et al. 2011

Biotech & Bioengineering

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The effect of ligand density on anion-exchange membrane chromatography (AEXmc) for the purification of recombinant baculoviruses (rBVs), potential viral vectors in clinical applications, is studied by surface plasmon resonance on customized AEX surfaces and gradient elution experiments on Sartobind D membrane prototypes with different diethylamine ligand densities, complemented by dynamic light scattering analysis for estimation of the hydrodynamic particle size of the various biologics. A chromatographic-column model based on the steric mass action model of ion exchange is employed to analyze the gradient-elution AEXmc experiments, extrapolate the results to other operating conditions, and provide directions for process improvement. Although counterintuitively, the experimental evidence provided in this study shows that the lowering of ligand density is beneficial for rBV purification by AEXmc in bind-and-elute-mode, because it decreases the residual concentrations of host cell protein, dsDNA, and non-infective rBVs in the eluted product cut, and increases the overall yield by roughly 20% over current standard values. Overall, we present a case study on how rational design can streamline downstream process development.

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“…Ion-exchange chromatography has also been used to purify viruses from cell debris based on the net surface charges of viruses (Aasted et al 1984;Slepushkin et al 2003;Smith 1987;Trilisky and Lenhoff 2007;Walin et al 1994). During ionexchange chromatography, charged particles are bound to the column matrix, with negatively charged particles replacing negatively charged ions on the column matrix for anionexchange chromatography, or positively charged particles replacing positively charged ions on the matrix for cationexchange chromatography (Scopes 1993).…”

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confidence: 99%

Physical fractionation of aquatic viral assemblages

Brum

Steward

2011

Limnology & Ocean Methods

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Aquatic viruses are extraordinarily diverse and have major influences on plankton ecology and evolution, but the majority of these viruses remain uncharacterized. Most viruses have not been cultivated and cultivationindependent means to explore viral diversity have provided only a fragmented view of viral genomic information. In this study, ultracentrifugation and chromatographic methods were evaluated for their ability to fractionate aquatic viruses based on their differing physical properties with the aim of physically enriching subsets of aquatic viruses for further study. Centrifugation in continuous cesium chloride gradients, strong and weak anion‐exchange chromatography, and size‐exclusion chromatography with Sephacryl S‐1000 were found to separate aquatic viruses based on their differing buoyant densities, surface charges, and sizes, respectively. Sucrose density gradients, cation‐exchange chromatography, and size‐exclusion chromatography using controlled‐pore glass were found to have insufficient resolution to effectively separate viral assemblages. Using the successful methods listed above, we were able to separate complex assemblages of viruses into fractions that had distinguishable subsets of the viruses present in the original community as determined by pulsed‐field gel electrophoresis. Strong anion‐exchange chromatography was particularly effective at separating viruses in the samples and resulted in one to six dominant viral genome bands in most fractions. Our results suggest that individual populations of viruses may be physically enriched, and possibly isolated, from complex assemblages without cultivation. These fractionation methods are expected to improve our ability to characterize the genotypes and phenotypes of the uncultivated majority of viruses in aquatic environments.

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Sorption processes in ion-exchange chromatography of viruses

Cited by 78 publications

References 45 publications

Adsorption of plasmid DNA on anion exchange chromatography media

Adsorption of plasmid DNA on anion exchange chromatography media

Impact of ligand density on the optimization of ion‐exchange membrane chromatography for viral vector purification

Physical fractionation of aquatic viral assemblages

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