Capillary electrophoresis of proteins 2005–2007

Dolnı́k, Vladislav

doi:10.1002/elps.200700584

Cited by 152 publications

(112 citation statements)

References 230 publications

(198 reference statements)

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“…Innovations involving both protocol formulations and new experimental setups are continuously proposed in the literature. In particular, relevant reviews including CZE of proteins and peptides are available (see, for instance, [2][3][4][5] and citations therein). At present CZE models are useful in the physicochemical characterization and interpretation of the effective mobility data of peptides and proteins [6][7][8][9][10][11][12][13][14][15][16].…”

Section: Introductionmentioning

confidence: 99%

Analysis of the interplay among charge, hydration and shape of proteins through the modeling of their CZE mobility data

2009

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Electrophoretic mobility data of four proteins are analyzed and interpreted through a physicochemical CZE model, which provides estimates of quantities like equivalent hydrodynamic radius (size), effective charge number, shape orientation factor, hydration, actual pK values of ionizing groups, and pH near molecule, among others. Protein friction coefficients are simulated through the creeping flow theory of prolate spheroidal particles. The modeling of the effective electrophoretic mobility of proteins requires consideration of hydrodynamic size and shape coupled to hydration and effective charge. The model proposed predicts native protein hydration within the range of values obtained experimentally from other techniques. Therefore, this model provides consistently other physicochemical properties such as average friction and diffusion coefficients and packing fractal dimension. As the pH varies from native conditions to those that are denaturing the protein, hydration and packing fractal dimension change substantially. Needs for further research are also discussed and proposed.

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Section: Introductionmentioning

confidence: 99%

Analysis of the interplay among charge, hydration and shape of proteins through the modeling of their CZE mobility data

2009

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“…CE has proved to be an effective method for protein analysis [1,2]. Besides its high separation efficiency, the advantages of CE include low sample consumption and short analysis time [3,4].…”

Section: Introductionmentioning

confidence: 99%

Hydroxyethylcellulose‐graft‐poly (4‐vinylpyridine) as a novel, adsorbed coating for protein separation by CE

2009

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A novel graft copolymer, hydroxyethylcellulose-graft-poly (4-vinylpyridine), used as a physically adsorbed coating of the fused-silica (FS) capillaries, was synthesized by using ceric ammonium nitrate initiator in aqueous nitric acid solution. EOF measurements showed that the above graft copolymer coating of the FS capillaries could suppress EOF effectively compared with the bare FS capillaries. The adsorbed coating exhibited minimal interactions with proteins, providing efficient protein separations with excellent repeatability. Interestingly, the electrical charge of the coated capillary wall could be modulated by varying the pH of the running buffer, which makes possible the analysis of basic and acidic proteins in the same capillary. The effects of pH on suppressing EOF and protein separation were investigated in detail. The results demonstrated that the hydroxyethylcellulose-graft-poly (4-vinylpyridine) copolymer coatings have great potential in the diagnosis and proteomics.

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“…After the optimization of pH, we also optimized the buffer species. Three kinds of buffer solutions, H 3 PO 4 À NaH 2 PO 4 buffer, CH 3 COOH-CH 3 COONa buffer and CH 3 COOH-Tris buffer, were used to examine the effect of buffer solution on separation efficiency. The experiments showed that the best sensitivity and resolution could be achieved when CH 3 COOH-Tris buffer was used as the running solution.…”

Section: Analytical Application For Protein Detectionmentioning

confidence: 99%

“…CE possesses the advantages of high resolution, rapid separation and ease of automation. Therefore, the great potential of CE has been envisioned for the analysis of proteins and investigation of proteomes [1][2][3]. However, a major challenge in CE detection of protein is the poor concentration detection limit due to the short optical path length across the capillary and the small sample volumes injected.…”

Section: Introductionmentioning

confidence: 99%

CE with a new electrochemiluminescent detection system for separation and detection of proteins labeled with tris(1,10‐phenanthroline) ruthenium(II)

Guo¹,

Qiu²,

Xue³

2009

Electrophoresis

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In this article, a CE with a new electrochemiluminescent (ECL) detection system was developed. A microfluidic ECL detection cell with less than 0.5 microL dead volumes was developed and used as detector for this system. A hydrofluoric acid-etched porous joint was made at 8 mm from the outlet of the separation capillary to isolate the CE high voltage from ECL detection. The proposed CE-ECL system was applied for separation and detection of some proteins labeled with tris(1,10-phenanthroline) ruthenium(II). High efficiency ECL-enhanced reagent, tripropylamine, was infused to the detection cell as coreactant by a micro-infusion system to obtain maximum and stable ECL signal. The performance of this setup was illustrated by the analysis of tris(1,10-phenanthroline) ruthenium(II)-labeled proteins. The background electrolyte for protein detection was 20 mM Tris-CH3COOH with 2.0% m/m PVP at pH 4.0. Under the optimal conditions, the corresponding LOD were 2.2 x 10(-10) M for HSA, 4.4 x 10(-10) M for casein (alpha-S1) and 5.1 x 10(-10) M for cytochrome c. The proposed method was also successfully used for the trace analysis of albumin in human urine without any pretreatment.

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Capillary electrophoresis of proteins 2005–2007

Cited by 152 publications

References 230 publications

Analysis of the interplay among charge, hydration and shape of proteins through the modeling of their CZE mobility data

Analysis of the interplay among charge, hydration and shape of proteins through the modeling of their CZE mobility data

Hydroxyethylcellulose‐graft‐poly (4‐vinylpyridine) as a novel, adsorbed coating for protein separation by CE

CE with a new electrochemiluminescent detection system for separation and detection of proteins labeled with tris(1,10‐phenanthroline) ruthenium(II)

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