Ion Exchange Chromatography

Karlsson, Evert; Hirsh, Irwin

doi:10.1002/9780470939932.ch4

Cited by 17 publications

(9 citation statements)

References 109 publications

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“…IEC is widely used for the separation of differentially charged molecules, which are concentrated during binding to the matrix/column and then collected in a purified, concentrated form (16). We chose amniotic fluid (AF) as a model of a complex biological fluid and compared the current protocol for isolation of amniotic fluid EVs (afEVs), based on ultracentrifugation and SDG separation (17,18), with our novel protocol combining ultracentrifugation and IEC.…”

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confidence: 99%

Ion-exchange chromatography purification of extracellular vesicles

et al. 2017

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Despite numerous studies, isolating pure preparations of extracellular vesicles (EVs) has proven challenging. Here, we compared ion-exchange chromatography (IEC) to the widely used sucrose density gradient (SDG) centrifugation method for the purification of EVs. EVs in bulk were isolated from pooled normal human amniotic fluid (AF) by differential centrifugation followed by IEC or sucrose density gradient separation. The purity of the isolated EVs was evaluated by electrophoresis and lectin blotting/immuno blotting to monitor the distribution of total proteins, different EVs markers, and selected N-glycans. Our data showed efficient separation of negatively charged EVs from other differently charged molecules, while comparative profiling of EVs using SDG centrifugation confirmed anion-exchange chromatography is advantageous for EV purification. Finally, although this IEC-based method was validated using AF, the approach should be readily applicable to isolation of EVs from other sources as well.

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confidence: 99%

Ion-exchange chromatography purification of extracellular vesicles

et al. 2017

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“…The reusability of ion-exchangers is often hampered by the efficiency of their sanitization (cleaning in place) 15 16 . Anion exchangers are more prone to strongly binding pigments and other difficult to remove colouring substances 17 18 . Cation exchangers are easier to maintain in this regard and the investigated mixed mode resin is similar in this manner.…”

Section: Resultsmentioning

confidence: 99%

Mixed-mode resins: taking shortcut in downstream processing of raw-starch digesting α-amylases

Lončar

Slavić

Vujčić

et al. 2015

Sci Rep

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Bacillus licheniformis 9945a α-amylase is known as a potent enzyme for raw starch hydrolysis. In this paper, a mixed mode Nuvia cPrime™ resin is examined with the aim to improve the downstream processing of raw starch digesting amylases and exploit the hydrophobic patches on their surface. This resin combines hydrophobic interactions with cation exchange groups and as such the presence of salt facilitates hydrophobic interactions while the ion-exchange groups enable proper selectivity. α-Amylase was produced using an optimized fed-batch approach in a defined media and significant overexpression of 1.2 g L−1 was achieved. This single step procedure enables simultaneous concentration, pigment removal as well as purification of amylase with yields of 96% directly from the fermentation broth.

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“…Matrix has an ion load opposite to that of the protein to be separated, and the affinity of the protein to the column is achieved with ionic ties. Proteins are separated from the column either by changing pH, concentration of ion salts or ionic strength of the buffer solution [8]. Positively charged ion- exchange matrices are called anion-exchange matrices, and adsorb negatively charged proteins.…”

Section: Ion- Exchange Chromatographymentioning

confidence: 99%

Separation Tecniques: CHROMATOGRAPHY

Coşkun

2016

North Clin Istanbul

149

116

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Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis. Proteins can be purified based on characteristics such as size and shape, total charge, hydrophobic groups present on the surface, and binding capacity with the stationary phase. Four separation techniques based on molecular characteristics and interaction type use mechanisms of ion exchange, surface adsorption, partition, and size exclusion. Other chromatography techniques are based on the stationary bed, including column, thin layer, and paper chromatography. Column chromatography is one of the most common methods of protein purification.

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Ion Exchange Chromatography

Cited by 17 publications

References 109 publications

Ion-exchange chromatography purification of extracellular vesicles

Ion-exchange chromatography purification of extracellular vesicles

Mixed-mode resins: taking shortcut in downstream processing of raw-starch digesting α-amylases

Separation Tecniques: CHROMATOGRAPHY

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