Surface modification based on Si‐O and Si‐C sublayers and a series of <i>N</i>‐substituted acrylamide top‐layers for capillary electrophoresis

Gelfi, Cecilia; Curcio, Mario; Righetti, Pier Giorgio; Sebastiano, Roberto; Citterio, Attilio; Ahmadzadeh, Hossein; Dovic̀hi, Norman J.

doi:10.1002/elps.1150191026

Cited by 93 publications

(76 citation statements)

References 16 publications

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“…If an uncoated or dynamically coated capillary became fouled, or otherwise needed to be cleaned, the following wash procedure was performed at 20 psi: 0.5 M potassium hydroxide for 10 min, 0.5 M hydrochloric acid for 5 min, water for 5 min, and CE (uncoated) or PVA (dynamic) buffer for 5 min. AAP coated capillaries were prepared as described previously [28]. The instrument was aligned using a continuous flow of 10 -9 M fluorescein in CE buffer.…”

Section: Capillary Electrophoresismentioning

confidence: 99%

Evaluation of individual particle capillary electrophoresis experiments via quantile analysis

Whiting

Arriaga

2007

Journal of Chromatography A

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The number of particles in a sample heavily influences the shape of the distribution describing the corresponding individual particle measurements. Selecting an adequate number of particles that prevents biases due to sample size is particularly difficult for complex biological systems in which statistical distributions are not normal. Quantile analysis is a powerful statistical technique that can rapidly compare differences between multiple distributions of individual particles. This report utilizes quantile analysis to show that the number of events detected affects the mobility distributions for rat liver and mouse liver mitochondria, sample individual particles, when analyzed via capillary electrophoresis with laser-induced fluorescence. When the mitochondrial sample is small (e.g. <78), there are not enough events to obtain statistically relevant mobility data. Adsorption to the capillary surface also significantly affects the mobility distribution at a small number of events in uncoated and dynamically coated capillaries. These adsorption effects can be overcome when the mitochondrial load on the capillary is sufficiently large (i.e. >609 and >1426 events for mouse liver on uncoated capillaries and rat liver on dynamically coated capillaries, respectively). It is anticipated that quantile analysis can be used to study other distributions of individual particles, such as nanoparticles, organelles, and biomolecules, and that distributions of these particles will also be dependant on sample size.

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Section: Capillary Electrophoresismentioning

confidence: 99%

Evaluation of individual particle capillary electrophoresis experiments via quantile analysis

Whiting

Arriaga

2007

Journal of Chromatography A

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“…In this experiment, we flush the capillary between runs to remove adsorbed proteins that might interfere with subsequent separations. We have demonstrated that modified acrylamide coatings are useful in reducing protein adsorption during capillary electrophoresis (21). Such coatings will be required for the analysis of highly basic and highly hydrophobic proteins, which were likely lost during our analyses.…”

Section: Discussionmentioning

confidence: 98%

Fully Automated Two-dimensional Capillary Electrophoresis for High Sensitivity Protein Analysis

Michels

Schoenherr

et al. 2002

Molecular & Cellular Proteomics

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117

114

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We report a system for automated protein analysis. In the system, proteins are labeled with the fluorogenic reagent 3-(2-furoyl)quinoline-2-carboxaldehyde, which reacts with lysine residues and creates a highly fluorescent product. These labeled proteins are analyzed by submicellar capillary electrophoresis at pH 7.5 to perform a first dimension separation. Once the first components migrate from the capillary, a fraction is transferred to a second dimension capillary, where electrophoresis is performed at pH 11.1 to further separate the proteins. Laser-induced fluorescence is used as an ultrasensitive detector of the separated proteins. Successive fractions are transferred from the first dimension capillary to the second dimension capillary for further separation to generate, in serial fashion, a twodimensional electropherogram. The transfer of fractions is computer-controlled; there is no operator intervention once the sample has been injected. Zeptomoles of labeled proteins are detected, providing exquisite sensitivity. The resolution of complex samples into components requires sophisticated technology. Most separation techniques are capable of resolving, at most, several dozen components. Cal Giddings (1) recognized that the combination of two separation techniques is important in the resolution of complex mixtures. If the two separation techniques are based on unrelated characteristics of the sample, then the number of resolution elements is given by the product of the resolution elements of both separation steps. For example, isoelectric focusing and SDS-polyacrylamide gel electrophoresis can, individually, resolve ϳ50 components in a protein sample. Their combination, in two-dimensional electrophoresis, can resolve several thousand components (2).Unfortunately, classic two-dimensional electrophoresis requires manual manipulation of the sample. These manipulations, although reasonable for occasional use, are very tedious when performed in large-scale protein analysis projects. Furthermore, detection sensitivity is limited, and relatively large amounts of sample are required to detect weakly expressed components.Column-switching technology is an alternative means of performing two-dimensional separations and is commonly used in chromatographic resolution of complex samples. Most simply, a fraction containing the compound of interest is captured as it elutes from the first column and is transferred to another chromatographic column for additional resolution; fractionation is repeated with different chromatographic methods to achieve the desired purity.In multidimensional chromatography, also known as comprehensive chromatography, a second column is used to sequentially separate all fractions from the first column. These two-dimensional techniques are particularly useful when characterizing extremely complex samples. An early report used two successive chromatographic steps to purify peptides generated from the proteolytic digest of a human immunoglobulin (3).Jorgenson and others (4 -12) have developed elegant...

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“…Gelfi et al [176] polymerized either acrylic acid (AA), DMA, N-acryloylaminoethoxyethanol (AAEE), or N-acryloylaminopropanol (AAP). A direct Si-C bond at the surface was formed by a silanol chlorination of the surface, followed by a Grignard coupling of a vinylmagnesium bromide [177] to increase resistance of the coating to alkaline hydrolysis.…”

Section: Polymer Brushes By Grafting To Strategymentioning

confidence: 99%

Surface treatment and characterization: Perspectives to electrophoresis and lab‐on‐chips

et al. 2006

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The control and modification of surface state is a major challenge in bioanalytical sciences, and in particular in electrokinetic separation methods, due to the importance of electroosmosis. This topic has gained recently a renewed interest, associated with the development of "lab-on-chips" systems that extend the range of materials in which separation channels are fabricated. The surface science community has developed through the years a large toolbox of characterization tools and surface modification protocols, which is not yet fully exploited in the bioanalytical world. In this paper, we try and present an overview of these tools, in order to stimulate new ideas for improved and more controlled surface treatment strategies for separations in capillaries and microchannels. We briefly describe some physical and chemical aspects of electroosmosis (global and spatially resolved), streaming current, and streaming potential. We also review the main strategies for surface coating, and compare the advantages of physisorption, well-organized thin self-assembled monolayers, or conversely thick polymer "brushes". Examples of existing applications to electrophoresis in microchannel are also given.

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Surface modification based on Si‐O and Si‐C sublayers and a series of N‐substituted acrylamide top‐layers for capillary electrophoresis

Cited by 93 publications

References 16 publications

Evaluation of individual particle capillary electrophoresis experiments via quantile analysis

Evaluation of individual particle capillary electrophoresis experiments via quantile analysis

Fully Automated Two-dimensional Capillary Electrophoresis for High Sensitivity Protein Analysis

Surface treatment and characterization: Perspectives to electrophoresis and lab‐on‐chips

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