Polyethylenimine modified poly(ethylene terephthalate) capillary channeled-polymer fibers for anion exchange chromatography of proteins

Jiang, Liuwei; Jin, Yi; Marcus, R. Kenneth

doi:10.1016/j.chroma.2015.07.102

Cited by 29 publications

(11 citation statements)

References 49 publications

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“…In addition, a physically coated PEI onto gold nanoparticles fixed in a capillary monolithic column was also used in HILIC mode [25]. More recently, PEI-modified polymer fibers as anion exchanger were used for separation of proteins [26]. Much similar to RPLC, the majority of HILIC SPs are based on silica gel since it has many well-documented advantages such as easy modification.…”

Section: Introductionmentioning

confidence: 99%

A hyperbranched polyethylenimine functionalized stationary phase for hydrophilic interaction liquid chromatography

et al. 2016

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A hyperbranched stationary phase for hydrophilic interaction liquid chromatography (HILIC) has been prepared by grafting polyethylenimine (PEI) onto silica gel (termed as PEI-Sil). Rich primary, secondary, and tertiary amino groups associated with PEI render its good hydrophility. More importantly, the hyperbranched structure of PEI molecule is greatly helpful in improving interaction with polar analytes. For several kinds of model polar compounds, including organic acids, nucleosides, nucleic acid bases, amino acids, cephalosporins, and non-reducing sugars, PEI-Sil demonstrated excellent separation performance in terms of running stability, reproducibility, and separation efficiency (e.g., plate count ~74,000/m). In addition, PEI-Sil also exhibited much better separation selectivity toward inorganic anions when operated in the mode of ion chromatography relative to a commercial amino propyl-bonded column.

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Section: Introductionmentioning

confidence: 99%

A hyperbranched polyethylenimine functionalized stationary phase for hydrophilic interaction liquid chromatography

et al. 2016

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“…These 30‐ to 50‐μm diameter polymer fibers are made in a melt‐spin process by extrusion from PP, PET, and nylon 6 base materials through an orifice with the desired shape. Fiber surfaces can be modified on‐column through simple adsorption or covalent ligand coupling to provide greater selectivity . The close, parallel packing of the eight‐channeled fibers provides a unique transport medium.…”

Section: Introductionmentioning

confidence: 99%

“…Marcus and co-workers have investigated the merits of capillary-channeled polymer (C-CP) fiber stationary phases 13,14,40 for protein separations under RP-LC, ion exchange chromatography and affinity chromatography [41][42][43][44][45][46][47] conditions. These 30-to 50-μm diameter polymer fibers are made in a melt-spin process by extrusion from PP, PET, and nylon 6 base materials through an orifice with the desired shape.…”

Section: Introductionmentioning

confidence: 99%

“…Fiber surfaces can be modified on-column through simple adsorption or covalent ligand coupling to provide greater selectivity. [41][42][43][45][46][47] The close, parallel packing of the eightchanneled fibers provides a unique transport medium. Very low intrafiber porosity, 48 allows protein separations at extremely high linear velocities (100 mm s −1 ) without negative effects of intraphase mass transfer (van Deemter C-term) broadening.…”

Section: Introductionmentioning

confidence: 99%

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Overload Effects in Reversed Phase Protein Separations using Capillary‐Channeled Polymer Fiber Columns

Wang

Marcus

2018

Biotechnology Progress

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In preparative chromatography, overloading effects are a fact of life. However, peak distortion is problematic in analytical scale chromatography, especially in the case of bio-macromolecule separations. Capillary-channeled polymer fibers have been employed for fast protein separations in reversed phase, ion exchange, and hydrophobic interaction liquid chromatography. Although the primary advantage of the phase is operation at high linear velocities (~100 mm s ) without van Deemter C-term limitations, the limited specific surface area (<5 m g ) suggests that a thorough understanding of overloading effects is needed. We evaluate important factors (injected mass and volume) affecting overload in terms of peak height, width and shape (asymmetry). Overload conditions readily apparent as the peak shape changes from a Gaussian distribution to a left-triangle with slight tailing; more-or-less classical overload behavior. Three methods were used to compute column efficiency in terms of plate counts (N), including the area height, half-height, and the Dorsey-Foley approaches. The half-height method is best-suited for describing column overload, accounting for peak distortion and achieving high levels of consistency. The limiting plate count (N ) and the column sample loading capacity (ω ) were key metrics to characterize overall column performance. Peak capacity (P) was used to assess gradient elution separation performance. Increased flow rates and column length result in enhanced loading capacity. pH mismatch between the sample matrix and the mobile phase, a common cause of protein overload with other phases, was not responsible for distorted peak shapes unless the solvent pH is close to the pI of protein samples.

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“…[25][26][27][28] Previous works have employed C-CP fibers extensively in intact protein separations via various modalities. [29][30][31][32][33] HIC is a mode of separation in which molecules (usually proteins) in a high salt concentration environment interact hydrophobically with the lesspolar surfaces of the stationary phase 34 ; a variant of the common "salting out" mode of protein purification. Elution is affected by reduction of the solvent salinity, resolvating the molecules into the eluate flow.…”

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confidence: 99%

Evaluation of exosome loading characteristics in their purification via a glycerol‐assisted hydrophobic interaction chromatography method on a polyester, capillary‐channeled polymer fiber phase

Huang

Wang

Bruce

et al. 2020

Biotechnology Progress

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Exosomes are membrane-secreted vesicles, with sizes ranging from 30 to 150 nm, which play key roles in intercellular communication. There is intense interest in developing methods to isolate and quantify exosomes toward clinical diagnostics, fundamental studies of intercellular processes, and use of exosomes as delivery vehicles for therapeutic agents. Current methods for exosomes isolation and quantification are time consuming and have operational high costs; few combine isolation and quantification into a singular operation unit. This report describes the use of hydrophobic interaction chromatography on a polyester capillary-channeled polymer fiber column, employing a step gradient for exosome elution, including use of glycerol as a solvent modifier. The entire procedure is completed in 8 min, while maintaining the structural integrity and biological activity of the isolated exosomes. Electron microscopy was used to verify the size and structural fidelity of single exosomes. Absorbance response curves for a commercial exosome sample were used for exosome quantification in the chromatographic separations. In order to determine the dynamic loading capacity for exosomes, different volumes of Dictyostelium discoideum cell culture milieu supernatant were loaded at different column lengths (5-30 cm) and loading flow rates (0.2-0.5 ml/min). A loading capacity of 5.4 × 10 12 exosomes derived from D. discoideum milieu was obtained on a 0.8 × 300 mm column; yielding recoveries of over 80%. It is believed that this isolation and purification strategy holds many advantages toward the use of exosomes across a wide breadth of medical and biotechnology applications. K E Y W O R D S capillary-channeled polymer fibers, exosomes, extracellular vesicles, hydrophobic interaction chromatography, purification 1 | INTRODUCTION Exosomes, a subset of extracellular vesicles (EVs), are membranesecreted vesicles, with sizes ranging from 30 to 150 nm, which exist in almost all cell types and body fluids and play key roles in intercellular communication and cancer progression. 1,2 Formed in the multivesicular body, exosomes encapsulate various species including proteins and genetic material, and carry along surface proteins representative of their source. Notably, tumor cells have been shown to produce and secrete exosomes in greater 3, 4 numbers than normal

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Polyethylenimine modified poly(ethylene terephthalate) capillary channeled-polymer fibers for anion exchange chromatography of proteins

Cited by 29 publications

References 49 publications

A hyperbranched polyethylenimine functionalized stationary phase for hydrophilic interaction liquid chromatography

A hyperbranched polyethylenimine functionalized stationary phase for hydrophilic interaction liquid chromatography

Overload Effects in Reversed Phase Protein Separations using Capillary‐Channeled Polymer Fiber Columns

Evaluation of exosome loading characteristics in their purification via a glycerol‐assisted hydrophobic interaction chromatography method on a polyester, capillary‐channeled polymer fiber phase

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