Field‐flow fractionation of proteins, polysaccharides, synthetic polymers, and supramolecular assemblies

Williams, S. Kim Ratanathanawongs; Lee, Dean

doi:10.1002/jssc.200600151

Cited by 125 publications

(29 citation statements)

References 125 publications

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“…Virus diameters typically range between 20 and 300 nm (King et al 2012) and thus they are expected to elute in normal mode. AF4 has been widely used for separation and characterisation of submicron-and micron-sized bioparticles including viruses [for reviews see (Ratanathanawongs and Williams 2006;Roda et al 2009)]. It has been exploited to optimise virus-like particle (VLP) production in bacterial, yeast and mammalian cells, to assay VLP assembly and disassembly conditions, to determine size distribution of sample components in virus and VLP specimens, to quantitate particle amounts, and to study the presence of aggregates and fragments in particle preparations (Chen et al 2015;Chuan et al 2008;Lang et al 2009;Somasundaram et al 2016;Wei et al 2007).…”

Section: Introductionmentioning

confidence: 99%

Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation

et al. 2017

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Viruses come in various shapes and sizes, and a number of viruses originate from extremities, e.g. high salinity or elevated temperature. One challenge for studying extreme viruses is to find efficient purification conditions where viruses maintain their infectivity. Asymmetrical flow field-flow fractionation (AF4) is a gentle native chromatography-like technique for size-based separation. It does not have solid stationary phase and the mobile phase composition is readily adjustable according to the sample needs. Due to the high separation power of specimens up to 50 µm, AF4 is suitable for virus purification. Here, we applied AF4 for extremophilic viruses representing four morphotypes: lemon-shaped, tailed and tailless icosahedral, as well as pleomorphic enveloped. AF4 was applied to input samples of different purity: crude supernatants of infected cultures, polyethylene glycol-precipitated viruses and viruses purified by ultracentrifugation. All four virus morphotypes were successfully purified by AF4. AF4 purification of culture supernatants or polyethylene glycol-precipitated viruses yielded high recoveries, and the purities were comparable to those obtained by the multistep ultracentrifugation purification methods. In addition, we also demonstrate that AF4 is a rapid monitoring tool for virus production in slowly growing host cells living in extreme conditions.

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Section: Introductionmentioning

confidence: 99%

Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation

et al. 2017

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“…Asymmetric Frit Inflow Field Flow Fractionation (FIFFF) and Thermal Field Flow Fractionation (ThFFF) are recently applied to correlate the size of prion particles with infectivity. Protease resistant PrP protein aggregates were fractioned and the molar mass and hydrodynamic radius was determined using Quasi Elastic light scattering (QELS) [27].…”

Section: Single Molecule Based Methodsmentioning

confidence: 99%

Molecular Nanobiotechnological Approaches for the Detection and Therapy of Prion Related Diseases

P¹,

Thampy²,

Venugopalan³

et al. 2012

Nano BioMed ENG

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Prion diseases are associated with the accumulation in the brain of an abnormal, protease resistant isoform of a host encoded glycoprotein known as prion protein (PrP). Nanotechnology in combination with biotechniques promises a broad spectrum of highly innovative approaches for overcoming the challenges posed by the prions. Recent advances in molecular nanobiotechnology have brought in the potential of molecular targeting in diagnosis and therapies of various diseases. Their high binding sensitivity and specificity added by their small size have favored the identification by in vitro protocols. Molecular targeting has initiated exciting technologies based on conjugation of biomolecules to nanoparticles. This review article is an extensive study of various research oriented nanobiotechnological protocols for rapid identification and cure for prion diseases both at in vivo and in vitro options.

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“…New materials are created every day, and it is just a matter of time when membranes with improved solvent resistance are going to be introduced for AF4. Thermal FFF uses a thermal gradient for separation and is mostly used for the analysis of lipophilic‐shear‐sensitive synthetic polymers or aggregates in both water and organic solvents . It was also shown to be applicable for the separation of some starches in DMSO, yet difficult for good characterization of branched polysaccharides .…”

Section: Oxymoron: Simply Hard To Characterize!mentioning

confidence: 99%

Polysaccharide Biocatalysis: From Synthesizing Carbohydrate Standards to Establishing Characterization Methods

Ćirić

Petrović

Loos

2014

Macro Chemistry & Physics

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Starch, glycogen, and cellulose are all around us. They are eaten and used on a daily basis but they are not understood completely. Even though these carbohydrates are simple, concerning their repeating unit, they are hard to characterize. In order to try to understand as much as possible about their structure and the relationship between their molecular structure and physical properties, it is very practical to create such polysaccharides, for instance enzymatically, characterize them, and use them as standards for the characterization of natural ones. Therefore, the main objective of this Trend article is to outline different enzymatic routes to such carbohydrates, possibilities for their characterization, and the characterization of natural ones.

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Field‐flow fractionation of proteins, polysaccharides, synthetic polymers, and supramolecular assemblies

Cited by 125 publications

References 125 publications

Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation

Halophilic viruses with varying biochemical and biophysical properties are amenable to purification with asymmetrical flow field-flow fractionation

Molecular Nanobiotechnological Approaches for the Detection and Therapy of Prion Related Diseases

Polysaccharide Biocatalysis: From Synthesizing Carbohydrate Standards to Establishing Characterization Methods

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