Application of capillary electrophoretic chips in protein profiling of plant extracts for identification of genetic modifications of maize

Poboży, Ewa; Filaber, Monika; Koc, Anna; García-Reyes, Juan F.

doi:10.1002/elps.201300103

Cited by 9 publications

(5 citation statements)

References 66 publications

(95 reference statements)

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“…It has been mentioned that some differences in protein profiles have been found for albumins and globulins fraction . Using CZE with low‐pH BGE, significant differences in albumin fraction for transgenic and nontransgenic maize varieties were found .…”

Section: Resultsmentioning

confidence: 99%

“…Significant qualitative and quantitative differences in CZE–UV profiling were found in the albumin fraction extracted from transgenic Aristis‐Bt and nontransgenic Aristis maize samples and in tryptic protein digests . Several repeatable differences were observed for total protein extracts of 0 and 5% m/m GMO maize standards using chip gel electrophoresis with LIF detection . On the other hand, CE–MS did not identify significant differences in the presence of the zein proteins composition of three GM maize lines (Aristis Bt, Tietar Bt, and PR33P66 Bt) and their corresponding control lines .…”

Section: Introductionmentioning

confidence: 93%

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Comparison of three modifications of fused‐silica capillaries and untreated capillaries for protein profiling of maize extracts by capillary electrophoresis

Poboży

Sentkowska

Piskor

2014

J of Separation Science

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In this work, capillary electrophoresis was applied to protein profiling of fractionated extracts of maize. A comparative study on the application of uncoated fused-silica capillaries and capillaries modified with hydroxypropylmethylcellulose, ω-iodoalkylammonium salt and a commercially available neutral capillary covalently coated with polyacrylamide is presented. The coating stability, background electrolyte composition, and separation efficiency were investigated. It was found that for zeins separation, the most stable and efficient was the capillary coated with polyacrylamide. Finally, the usefulness of these methods was studied for the differentiation of zein fraction in transgenic and nontransgenic maize. Zeins extracted from maize standards containing 0 and 5% m/m genetic modification were successfully separated, but slight differences were observed in terms of the zein content. Albumin and globulin fractions were analyzed with the use of unmodified fused-silica capillary with borate buffer pH 9 and the capillary coated with polyacrylamide with phosphate buffer pH 3. In the albumin fraction, additional peaks were found in genetically modified samples.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 93%

Comparison of three modifications of fused‐silica capillaries and untreated capillaries for protein profiling of maize extracts by capillary electrophoresis

Poboży

Sentkowska

Piskor

2014

J of Separation Science

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“…To control EOF, commercial coating agent (EOTrol TM , Target Discovery, USA) 10% v/v was added to the BGE composed of 5 mM sodium borate, pH 8.5, 3.5 mM SDS, 10% dextran m/v. Microchip format for SDS‐PAGE of proteins is commercially available from Agilent company (Waldbronn, Germany) . Several other examples of free ZE and GE separations of peptides and proteins are presented in Table .…”

Section: Electroseparation Modesmentioning

confidence: 99%

“…For protein profiling of different extracts of maize standards, GE with LIF detection was employed . For sensitivity enhancement, lyophilization, or preconcentration with cutoff filters were applied.…”

Section: Applicationsmentioning

confidence: 99%

Analysis of proteins and peptides by electromigration methods in microchips

Štěpánová

Kašička

2016

J of Separation Science

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This review presents the developments and applications of microchip electromigration methods in the separation and analysis of peptides and proteins in the period 2011-mid-2016. The developments in sample preparation and preconcentration, microchannel material, and surface treatment are described. Separations by various microchip electromigration methods (zone electrophoresis in free and sieving media, affinity electrophoresis, isotachophoresis, isoelectric focusing, electrokinetic chromatography, and electrochromatography) are demonstrated. Advances in detection methods are reported and novel applications in the areas of proteomics and peptidomics, quality control of peptide and protein pharmaceuticals, analysis of proteins and peptides in biomatrices, and determination of physicochemical parameters are shown.

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“…Protein profiles obtained by chip-electrophoresis have also been employed to investigate GMOs in maize samples [ 56 ]. For this purpose, albumin, globulin and zein fractions were analyzed using two gel chips in parallel with different mass ranges (5–80 and 14–230 kDa).…”

Section: Applications Of Chip-based Electrophoresis To the Study Amentioning

confidence: 99%

Characterization and Study of Transgenic Cultivars by Capillary and Microchip Electrophoresis

Vega

Marina

2014

IJMS

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Advances in biotechnology have increased the demand for suitable analytical techniques for the analysis of genetically modified organisms. Study of the substantial equivalence, discrimination between transgenic and non-transgenic cultivars, study of the unintended effects caused by a genetic modification or their response to diverse situations or stress conditions (e.g., environmental, climatic, infections) are some of the concerns that need to be addressed. Capillary electrophoresis (CE) is emerging as an alternative to conventional techniques for the study and characterization of genetically modified organisms. This article reviews the most recent applications of CE for the analysis and characterization of transgenic cultivars in the last five years. Different strategies have been described depending on the level analyzed (DNA, proteins or metabolites). Capillary gel electrophoresis (CGE) has shown to be particularly useful for the analysis of DNA fragments amplified by PCR. Metabolites and proteins have been mainly separated using capillary zone electrophoresis (CZE) using UV and MS detection. Electrophoretic chips have also proven their ability in the analysis of transgenic cultivars and a section describing the new applications is also included.

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Application of capillary electrophoretic chips in protein profiling of plant extracts for identification of genetic modifications of maize

Cited by 9 publications

References 66 publications

Comparison of three modifications of fused‐silica capillaries and untreated capillaries for protein profiling of maize extracts by capillary electrophoresis

Comparison of three modifications of fused‐silica capillaries and untreated capillaries for protein profiling of maize extracts by capillary electrophoresis

Analysis of proteins and peptides by electromigration methods in microchips

Characterization and Study of Transgenic Cultivars by Capillary and Microchip Electrophoresis

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