Capillary electrophoresis of peptides

Kašička, Václav

doi:10.1002/(sici)1522-2683(19991001)20:15/16<3084::aid-elps3084>3.0.co;2-4

Cited by 121 publications

(58 citation statements)

References 241 publications

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“…Most current CZE applications for peptides have involved relatively simple model mixtures or enzymatic digests of individual proteins [16,19]. Our previous comparison between CZE and gradient capillary reversed-phase liquid chromatography (RPLC) in the separation of tryptic digests of yeast lysate showed that CZE yielded a lower resolution than the RPLC approach [37].…”

Section: Czementioning

confidence: 99%

“…For proteins, the titration curves are steep enough to focus proteins into a narrow zone, and it has been estimated that a peak capacity of ,100 can be achieved in gels [24]. For peptides, separations may be limited because the effective charge of peptides approaches zero over a broad pH range rather than within a narrow range (as for proteins) [16]. However, the zone length formed in isoelectric focusing depends on a narrow pH zone around the pI [41], and relatively flat slopes at the pH zone close to the pI are found even for proteins [42].…”

Section: Cief Resolving Power Sample Concentration and Sample Loadingmentioning

confidence: 99%

“…Liquid-phase separations, often within capillaries, are increasingly recognized as the best separations technique for this approach. Developments in capillary electrophoresis applicable to separating peptides and proteins have been reviewed in several Electrophoresis publications [16][17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

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Proteomics based on high-efficiency capillary separations

2002

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Identifying and quantifying in a high throughput manner the proteins expressed by cells, tissues or an organism provides the basis for understanding the functions of its constituents at a "systems" level. As a result, proteome analysis has increasingly become the focus of significant interest and research over the past decade. This is especially true following the recent stunning achievements in genomics analyses. However, unlike the static genome, the complexities and dynamism of the proteome present significant analytical challenges and demand highly efficient separations and detection technologies. A number of recent technological advancements have been in direct response to these challenges. Currently, strategically mated combinations of sophisticated separations techniques and advanced mass spectrometric detection represent the best approach to addressing the intricacies of the proteome. Liquid-phase separations, often within capillaries, are increasingly recognized as the best separations technique for this approach. In combination on-line with mass spectrometry, liquid-phase separations provide the improved analytical sensitivity, sample throughput, and quantitation capabilities necessitated by the multifaceted problems within proteomics analyses. This review focuses primarily on current high-efficiency capillary separations techniques, including both capillary liquid chromatography and capillary electrophoresis, applied to the analysis of complex proteomic samples. We emphasize developments at our laboratory and illustrate technical advances that attempt to review the role of separations within the broader context of a state-of-the-art integrated proteomics effort.

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Section: Czementioning

confidence: 99%

Section: Cief Resolving Power Sample Concentration and Sample Loadingmentioning

confidence: 99%

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Proteomics based on high-efficiency capillary separations

2002

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“…Both large and small peptides can be separated by capillary electrophoresis (CE) [19]. In fact, because the basis of the separation is not at all related to hydrophobicity, CE is, in many ways, ideal for separating peptide mixtures that may consist of at least some large (> 20 aa) peptides [20].…”

Section: Separation Of Neuropeptidesmentioning

confidence: 99%

Techniques for neuropeptide determination

Sandberg

Weber

2003

TrAC Trends in Analytical Chemistry

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Because immunoassay responds to epitopes, and many molecules share the same peptide epitope, it is very difficult to obtain an accurate understanding of peptides, their creation and hydrolysis, in biological systems. Separate-and-detect approaches have merit in that the many active peptides and inactive fragments of a particular system can be separately determined. This review discusses the separation, by chromatography and capillary electrophoresis, and detection, by absorbance, fluorescence, electrochemistry, and immunoassay techniques. When separation pre-concentration is accompanied by laser-induced fluorescence or biuret-based electrochemical detection, nM-pM detection limits are obtained.

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“…CE is also a good separation technique for routine analysis of peptides [147]. CZE and MEKC separate large peptides molecules in a complex mixture.…”

Section: Ce-lifd Coupled To Microdialysis Studies Neuroactive Substamentioning

confidence: 99%

Biomedical applications of capillary electrophoresis with laser‐induced fluorescence detection

Páez

Hernández

2001

Biopharm & Drug Disp

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Capillary electrophoresis (CE) is a high-efficiency analytical technique that has had a great impact as a tool in biomedical research, clinical and forensic practice in the last ten years. Only in one of the applications, the DNA analysis, it has had an explosive exponential growth in the last few years. This impact is expressed in an enormous amount of CE articles and many reviews. The CE advantages with respect to other analytical techniques: the required very small sample volume, rapid analysis, great resolution power and low costs, have made this technique ideal for the analysis of a numerous endogenous and exogenous substances present in biological fluids. The different modes of CE have been coupled to different detection techniques such as UV-absorbance, electrochemical, mass spectrometry and laser-induced fluorescence detection (LIFD) to detect different nature and molecular size separated analytes. This review focuses mostly on the applications of CE-LIFD, to measure drugs and endogenous neuroactive substances such as amino acids and monoamines, especially in microdialysis samples from experimental animals and humans. CE-LIFD trends are discussed: automated faster analysis with capillary array systems, resolution power improvement, higher detection sensitivity, and CE systems miniaturization for extremely small sample volume, in order to make CE easier and affordable to the lab bench or the clinical bed.

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Capillary electrophoresis of peptides

Cited by 121 publications

References 241 publications

Proteomics based on high-efficiency capillary separations

Proteomics based on high-efficiency capillary separations

Techniques for neuropeptide determination

Biomedical applications of capillary electrophoresis with laser‐induced fluorescence detection

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