Chip‐Based Free‐Flow Electrophoresis with Integrated Nanospray Mass‐Spectrometry

Benz, Christian; Boomhoff, Michael; Appun, Johannes; Schneider, Christoph; Belder, Detlev

doi:10.1002/anie.201409663

Cited by 57 publications

(57 citation statements)

References 91 publications

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“…Various FFE modes, particularly the FF‐ZE and FF‐IEF have been used for prefractionation of complex peptide/protein mixtures in proteomic studies of cells and body fluids . FFE systems have been designed and applied to separation of peptides and proteins also in the microchip formats . However, due to their limited preparative capacity they are more suitable for continuous monitoring of selected analytes than for really preparative purposes.…”

Section: Applicationsmentioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2013–middle 2015)

Kašička

2015

Electrophoresis

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The review brings a comprehensive survey of recent developments and applications of high performance capillary and microchip electroseparation methods (zone electrophoresis, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography) to analysis, micropreparation, purification, and physicochemical and biochemical characterization of peptides in the years 2013, 2014, and ca. up to the middle of 2015. Advances in the investigation of electromigration properties of peptides, in the methodology of their analysis, including sample preseparation, preconcentration and derivatization, adsorption suppression and EOF control, as well as in detection of peptides, are described. New developments in particular CE and CEC modes are presented and several types of their applications to peptide analysis are reported: conventional qualitative and quantitative analysis, determination in complex (bio)matrices, monitoring of chemical and enzymatical reactions and physical changes, amino acid, sequence, and chiral analysis, and peptide mapping of proteins. Some 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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Section: Applicationsmentioning

confidence: 99%

Recent developments in capillary and microchip electroseparations of peptides (2013–middle 2015)

Kašička

2015

Electrophoresis

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“…In addition, miniaturization of FFZE methods on the microchip format can further allow significant savings in sample consumption and separation time [27‐30] besides rendering them readily integrable to other analytical procedures, for example, capillary zone electrophoresis [31] and sample preconcentration [32]. More recently, microchip‐based FFZE assays have been integrated to ESI‐MS systems by our research group [33] as well as others [34] allowing label‐free characterization of target analytes with high sensitivity. It is important to point out though that unlike other researchers, the FFZE platforms employed by our group for this application have relied on the use of internal electrodes placed across the width of the analysis channel.…”

Section: Introductionmentioning

confidence: 99%

Enhancement in MS‐based peptide detection by microfluidic free‐flow zone electrophoresis

2020

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Enhancement in MS-based peptide detection by microfluidic free-flow zone electrophoresisMatrix components are known to significantly alter the ionization of a target analyte in ESIbased measurements particularly when working with complex biological samples. This issue however may be alleviated by extracting the analyte of interest from the original sample into a relatively simple matrix compatible with ESI mass-spectrometric analysis. In this article, we report a microfluidic device that enables such extraction of small peptide molecules into an ESI-compatible solvent stream significantly improving both the sensitivity and reproducibility of the measurements. The reported device realizes this analyte extraction capability based on the free-flow zone electrophoretic fractionation process using a set of internal electrodes placed across the width of the analysis channel. Employing lateral electric fields and separation distances of 75 V/cm and 600 µm, respectively, efficient extraction of the model peptide human angiotensin II was demonstrated allowing a reduction in its detection limit by one to three orders of magnitude using the ESI-MS method. The noted result was obtained in our experiments both for a relatively simple specimen comprising DNA strands and angiotensin II as well as for human serum samples spiked with the same model peptide. Abbreviations: FFZE, free-flow zone electrophoresis; LLE, liquid-liquid extraction Color online: See article online to view Fig. 1-5 in color.

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“…Benz et al. presented a solution for coupling of μFFE outlet streams with electrospray ionization mass spectrometry (ESI‐MS) .…”

Section: Introductionmentioning

confidence: 99%

Micro free‐flow isoelectric focusing with integrated optical pH sensors

Nagl

2017

Engineering in Life Sciences

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Recently, a new observation method for monitoring of pH gradients in microfluidic free‐flow electrophoresis has emerged. It is based on the use of chip‐integrated fluorescent or luminescent micro sensor layers. These are able to monitor pH gradients in miniaturized separations in real time and spatially resolved; this is particularly useful in isoelectric focusing. Here these multifunctional microdevices that feature continuous separation, monitoring, and in some instances other functionalities, are reviewed. The employed microfabrication procedures to produce these devices are discussed and the different pH sensor matrices that were integrated and their applications in the separation of different types of biomolecules. The procedures for obtaining spatially resolved information about the separated molecules and the pH at the same time and different detection modalities to achieve this such as deep UV fluorescence as well as time‐resolved referenced pH sensing and the integration of a precolumn labeling step into these platforms are also highlighted.

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Chip‐Based Free‐Flow Electrophoresis with Integrated Nanospray Mass‐Spectrometry

Cited by 57 publications

References 91 publications

Recent developments in capillary and microchip electroseparations of peptides (2013–middle 2015)

Recent developments in capillary and microchip electroseparations of peptides (2013–middle 2015)

Enhancement in MS‐based peptide detection by microfluidic free‐flow zone electrophoresis

Micro free‐flow isoelectric focusing with integrated optical pH sensors

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