Separation of multiphosphorylated peptide isomers by CZE

Muetzelburg, Marika V.; Hoffmann, Ralf

doi:10.1002/elps.200800089

Cited by 14 publications

(16 citation statements)

References 28 publications

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“…As shown in Fig. 3, a full separation of all peptides was achieved in less than 14 min with a satisfactory resolution between P-T231/NP-T231 and P-S262/NP-S262 (4.8 and 8.7, respectively), similar to what was previously reported by Muetzelburg and Hoffmann [30].…”

Section: Resultssupporting

confidence: 86%

“…Furthermore, Li et al reported the analysis of phosphoproteins on an integrated glass chip that included a CE separation channel and MS detection [29]. Muetzelburg and Hoffmann have developed recently a CZE method using formic acid as BGE for the analysis of synthetic phosphopeptides related to tau protein [30].Tau is a normal brain phosphoprotein that belongs to the family of microtubule-associated proteins. Hyperphosphorylated forms of tau are the major constituents of the neurofibrillary lesions that are observed in Alzheimer's disease and other tauopathies.…”

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confidence: 99%

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A quantitative CE method to analyse tau protein isoforms using coated fused silica capillaries

Bohoyo-Gil

Potier

Klafki

et al. 2010

J of Separation Science

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We evaluated the potential of CE to analyse different isoforms of unphosphorylated recombinant tau protein and for separating one phosphorylated tau from the respective unphosphorylated protein. Different capillary coatings such as polyacrylamide, poly-(ethylene oxide) and polybrene (PB) were evaluated to overcome the poor efficiencies obtained with fused-silica capillary. Although peak asymmetry values were quite similar for the three investigated coatings, the peak efficiencies were 35-fold and 5-fold higher with PB coating than with polyacrylamide and poly(ethylene oxide) coatings, respectively. The recovery percentage (over 97%) was satisfactory and confirmed the efficacy of PB coating to limit the adsorption of tau protein to capillary walls. Moreover, PB coating produced higher repeatability for migration times (RSD values <1.2%) in comparison to the neutral coatings. The potential of PB-modified capillary in producing high resolutive separations of one phosphorylated tau isoform from its unphosphorylated counterpart and of a mixture of phosphorylated and unphosphorylated tau peptides was demonstrated with 50 mM phosphate buffer pH 3.0. The separation of unphosphorylated tau isoform 352 (Tau-352) from Tau-352 phosphorylated in vitro by the mitogen-activated protein kinase ERK2, was accomplished in less than 15 min.

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Section: Resultssupporting

confidence: 86%

mentioning

confidence: 99%

A quantitative CE method to analyse tau protein isoforms using coated fused silica capillaries

Bohoyo-Gil

Potier

Klafki

et al. 2010

J of Separation Science

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“…Separation of both enantiomers and diastereomers of di‐, tri‐, and tetrapeptides was achieved by CE in 0.5 M acetic acid BGE using diMe‐β‐CD, (+)‐(18‐crown‐6)‐2,3,11,12,‐tetracarboxylic acid, combination of these two compounds, or L‐histidine as chiral selectors 185. Complete or at least partial separation of positional isomers of mono, doubly and triply phosphorylated peptide fragments of human tau protein was achieved by CZE in BGE composed of 3.9 mol/L formic acid and 30% v/v trifluoroethanol 253.…”

Section: Applicationsmentioning

confidence: 99%

Recent advances in CE and CEC of peptides (2007–2009)

Kašička

2009

Electrophoresis

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The review presents a comprehensive survey of recent developments of high-performance capillary electromigration methods, zone electrophoresis, ITP, IEF, affinity electrophoresis, EKC and electrochromatography, and their application to analysis, preparation and physicochemical characterization of peptides. New approaches to the theoretical description and experimental investigation of electromigration properties of peptides, and to methodology of their separations, such as sample preparation, adsorption suppression, EOF control and detection, are described. New developments in particular CE and CEC modes are reported and several types of their applications to peptide analysis are presented: conventional qualitative and quantitative analysis, determination in biomatrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid and sequence analysis, and peptide mapping of proteins. Few examples of micropreparative peptide separations are shown and capabilities of CE and CEC techniques to provide important physicochemical characteristics of peptides are demonstrated.

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“…Therefore, analysis of the change of the phosphorylation of endogenous peptides is valuable for the diagnosis of diseases. However, due to their low abundance and lower ionization efficiency than nonphosphorylated peptides, enrichment method is normally required for phosphopeptides analysis []. Recently, titanium ion‐immobilized mesoporous silica particles have been successfully used to enrich the endogenous phosphopeptides from human serum because of their high adsorption specificity toward phosphopeptides [].…”

Section: Introductionmentioning

confidence: 99%

Titanium‐containing magnetic mesoporous silica spheres: Effective enrichment of peptides and simultaneous separation of nonphosphopeptides and phosphopeptides

Zhu

et al. 2012

J of Separation Science

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Protein phosphorylation is a common posttranslational modification, and involved in many cellular processes. Like endogenous peptides, endogenous phosphopeptides contain many biomarkers of preclinical screening and disease diagnosis. In this work, titanium-containing magnetic mesoporous silica spheres were synthesized and applied for effective enrichment of peptides from both tryptic digests of standard proteins and human serum. Besides, the enriched peptides can be further separated into nonphosphopeptides and phosphopeptides by a simple elution. First, titanium-containing magnetic mesoporous silica spheres were synthesized by a sol-gel method and found to have high surface area, narrow pore size distribution, and useful magnetic responsivity. Then, as the prepared material was used for selective capturing of phosphopeptides, it demonstrated to have higher selectivity than commercial titanium dioxide. Moreover, via combination of size-exclusion mechanism, hydrophobic interaction, and affinity chromatography, titanium-containing magnetic mesoporous silica spheres were successfully applied to simultaneously extract and separate nonphosphopeptides and phosphopeptides from standard protein digestion and human serum.

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Separation of multiphosphorylated peptide isomers by CZE

Cited by 14 publications

References 28 publications

A quantitative CE method to analyse tau protein isoforms using coated fused silica capillaries

A quantitative CE method to analyse tau protein isoforms using coated fused silica capillaries

Recent advances in CE and CEC of peptides (2007–2009)

Titanium‐containing magnetic mesoporous silica spheres: Effective enrichment of peptides and simultaneous separation of nonphosphopeptides and phosphopeptides

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