Recent developments in CE and CEC of peptides

Kašička, Václav

doi:10.1002/elps.200700550

Cited by 118 publications

(92 citation statements)

References 252 publications

(301 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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“…In recent years CEC has gained importance as a micro high-performance liquid separation technique for peptides and proteins [1][2][3][4][5]. CEC, which is often described as a hybrid of CZE and LC [6,7], has demonstrated higher separation efficiencies and flow permeabilities than capillary HPLC [8][9][10][11][12][13][14][15][16][17][18][19][20][21][22].…”

Section: Introductionmentioning

confidence: 99%

Electrochromatographic retention of peptides on strong cation‐exchange stationary phases

Nischang¹,

Höltzel²,

Tallarek³

2010

Electrophoresis

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We analyze the systematic and substantial electrical field-dependence of electrochromatographic retention for four counterionic peptides ([Met5]enkephalin, oxytocin, [Arg8]vasopressin, and luteinizing hormone releasing hormone (LHRH) ) on a strong cation-exchange (SCX) stationary phase. Our experiments show that retention behavior in the studied system depends on the charge-selectivity of the stationary phase particles, the applied voltage, and the peptides' net charge. Retention factors of twice positively charged peptides ([Arg8]vasopressin and LHRH at pH 2.7) decrease with increasing applied voltage, whereas lower charged peptides (oxytocin and [Met5]enkephalin at pH 2.7, [Arg8]vasopressin and LHRH at pH 7.0) show a concomitant increase in their retention factors. The observed behavior is explained on the basis of electrical fieldinduced concentration polarization (CP) that develops around the SCX particles of the packing. The intraparticle concentration of charged species (buffer ions, peptides) increases with increasing applied voltage due to diffusive backflux from the enriched CP zone associated with each SCX particle. For twice charged and on the SCX phase strongly retained peptides the local increase in mobile phase ionic strength reduces the electrostatic interactions with the stationary phase, which explains the decrease of retention factors with increasing applied voltage and CP intensity. Lower charged and weaker retained peptides experience a much stronger relative intraparticle enrichment than the twice-charged peptides, which results in a net increase of retention factors with increasing applied voltage. The CP-related contribution to electrochromatographic retention of peptides on the SCX stationary phase is modulated by the applied voltage, the mobile phase ionic strength, and the peptides' net charge and could be used for selectivity tuning in difficult separations.

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“…In CE, proteins are typically detected using UV absorption, laser induced fluorescence (LIF), or by coupling to a mass spectrometer (MS) (Fonslow & Yates, 2009;Garcia-Campana, Taverna, & Fabre, 2007;Herrero, Ibanez, & Cifuentes, 2008;Kasicka, 2008;Stutz, 2005). A limitation when using UV detection is that the limit of detection (LOD) is in the micromolar range.…”

Section: Using Capillary Electrophoresis To Identify the Proteome Of mentioning

confidence: 99%

Proteomics Analysis of Kinetically Stable Proteins

Xia¹,

Manning²,

Zhang³

et al. 2012

Integrative Proteomics

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Recent developments in CE and CEC of peptides

Cited by 118 publications

References 252 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

Electrochromatographic retention of peptides on strong cation‐exchange stationary phases

Proteomics Analysis of Kinetically Stable Proteins

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