Improving separation optimization in capillary electrophoresis by using a general quality criterion

Peró-Gascón, Roger; Tascón, Marcos; Sanz‐Nebot, Victoria; Gagliardi, Leonardo Gabriel; Benavente, Fernando

doi:10.1016/j.talanta.2019.120399

Cited by 5 publications

(4 citation statements)

References 28 publications

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“…Various aspects of the BGE selection for CE separations of peptides and proteins, such as type and concentration of the BGE constituents with respect to their buffering capacity, mobility and electric conductivity, pH, additives suppressing sorption of peptides/proteins to the inner surface of the most often used fused silica (FS) capillaries or influencing separation selectivity, EOF direction and velocity, organic modifiers, temperature, and Joule's heating effects, were in detail described in the earlier published but still relevant articles [48][49][50]. Useful information and practical advices for CZE and CIEF analysis of peptides and proteins can be found also in the tutorial [51] and in the articles dealing with multivariate techniques [52] and general quality criteria for optimization of CE methods [53]. Suitable and unsuitable pH buffers in biological, biochemical, and environmental studies and their interactions with metal ions are described in [54].…”

Section: Electromigration Properties Of Peptides and Selection Of Sep...mentioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2019–mid 2021)

Kašička

2021

Electrophoresis

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The review provides a comprehensive overview of developments and applications of high performance capillary and microchip electroseparation methods (zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) for analysis, microscale isolation, and physicochemical characterization of peptides from 2019 up to approximately the middle of 2021. Advances in the investigation of electromigration properties of peptides and in the methodology of their analysis, such as sample preparation, sorption suppression, EOF control, and detection, are presented. New developments in the individual CE and CEC methods are demonstrated and several types of their applications are shown. They include qualitative and quantitative analysis, determination in complex biomatrices, monitoring of chemical and enzymatic reactions and physicochemical changes, amino acid, sequence, and chiral analyses, and peptide mapping of proteins. In addition, micropreparative separations and determination of significant physicochemical parameters of peptides by CE and CEC methods are described.

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Section: Electromigration Properties Of Peptides and Selection Of Sep...mentioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2019–mid 2021)

Kašička

2021

Electrophoresis

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“…Useful information and practical advice for CZE and CIEF analysis of peptides and proteins can be found also in the tutorial [112] and in the papers dealing with multivariate techniques [148] and general quality criteria for the optimization of CE methods [149]. Suitable and unsuitable pH buffers in biological, biochemical, and environmental studies and their interactions with metal ions are described in [150].…”

Section: Separation Of Peptides From Digested Proteins By Ce Methodsmentioning

confidence: 99%

Peptide mapping of proteins by capillary electromigration methods

Kašička

2022

J of Separation Science

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This review article provides a wide overview of important developments and applications of capillary electromigration methods in the area of peptide mapping of proteins in the period 1997-mid-2022, including review articles on this topic. It deals with all major aspects of peptide mapping by capillary electromigration methods: i) precleavage sample preparation involving purification, preconcentration, denaturation, reduction and alkylation of protein(s) to be analyzed, ii) generation of peptide fragments by off-line or on-line enzymatic and/or chemical cleavage of protein(s), iii) postcleavage preparation of the generated peptide mixture for capillary electromigration separation, iv) separation of the complex peptide mixtures by one-, two-and multidimensional capillary electromigration methods coupled with mass spectrometry detection, and v) a large application of peptide mapping for variable purposes, such as qualitative analysis of monoclonal antibodies and other protein biopharmaceuticals, monitoring of posttranslational modifications, determination of primary structure and investigation of function of proteins in biochemical and clinical research, characterization of proteins of variable origin as well as for protein and peptide identification in proteomic and peptidomic studies.

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“…In previous works, the acid-base properties of opioid peptides [28] and Aβ peptide fragments [29] were characterized by CE-UV to establish electrophoretic migration models and systematically optimize their separation. Accurate pK a values were determined, which can be used to easily obtain charge-to-mass ratios (q/M r ) of the peptides and to calculate the multicriterium optimization function T ' for a simple, experiment-free and reliable selection of optimized separation conditions [30]. Figure 2A shows the plot of T ' (q/M r 1/2 ) vs pH for the mixture of the three opioid peptides (Dyn A, End 1, and Met) where a maximum indicates that the best separation will be obtained at pH values ranging from 3.0 to 3.5.…”

Section: Ce-msmentioning

confidence: 99%

Evaluation of on-line solid-phase extraction capillary electrophoresis–mass spectrometry with a nanoliter valve for the analysis of peptide biomarkers

Peró-Gascón

Benavente

Neusüß

et al. 2020

Analytica Chimica Acta

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On-line solid-phase extraction capillary electrophoresis-mass spectrometry (SPE-CE-MS) is a powerful technique for high throughput sample clean-up and analyte preconcentration, separation, detection, and characterization. The most typical design due to its simplicity and low cost is unidirectional SPE-CE-MS. However, in this configuration, the sample volumes introduced by pressure depend on the dimensions of the separation capillary and some matrix components could be irreversibly adsorbed in its inner walls. Furthermore, in many cases, the requirements of on-line preconcentration are incompatible with the background electrolyte necessary for an efficient separation and sensitive MS detection. Here, we present SPE-CE-MS with a nanoliter valve (nvSPE-CE-MS) to overcome these drawbacks while keeping the design simple. The nvSPE-CE-MS system is operated with a single CE instrument and two capillaries for independent and orthogonal SPE preconcentration and CE separation, which are interfaced through an external and electrically isolated valve with a 20 nL sample loop. The instrumental setup is proved for the analysis of opioid and amyloid beta peptide biomarkers in standards and plasma samples. NvSPE-CE-MS allowed decreasing the limits of detection (LODs) 200 times with regard to CE-MS. Compared to unidirectional SPE-CE-MS, peak efficiencies were better and repeatabilities similar, but total analysis times longer and LODs for standards slightly higher due to the heart-cut operation and the limited volume of the valve loop. This small difference on the LODs for standards was compensated for plasma samples by the improved tolerance of nvSPE-CE-MS to complex sample matrices. In view of these results, the presented setup can be regarded as a promising versatile alternative to avoid complicated matrix samples entering the separation capillary in SPE-CE-MS.

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Improving separation optimization in capillary electrophoresis by using a general quality criterion

Cited by 5 publications

References 28 publications

Recent developments in capillary and microchip electroseparations of peptides (2019–mid 2021)

Recent developments in capillary and microchip electroseparations of peptides (2019–mid 2021)

Peptide mapping of proteins by capillary electromigration methods

Evaluation of on-line solid-phase extraction capillary electrophoresis–mass spectrometry with a nanoliter valve for the analysis of peptide biomarkers

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