Novel polyamine coating providing non-covalent deactivation and reversed electroosmotic flow of fused-silica capillaries for capillary electrophoresis

Hardenborg, Emilia; Zuberovic, Aida; Ullsten, Sara; Söderberg, Linda; Heldin, Eva; Markides, Karin E.

doi:10.1016/s0021-9673(03)00739-8

Cited by 75 publications

(101 citation statements)

References 31 publications

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“…At pH 4.6, proteins were separated in less than 9 min with sufficient efficiencies ranging from 186 000 to 282 000 plates/m ( Table 2). These values were similar or superior to those obtained by capillaries coated with other cationic polymer [28,29].…”

Section: Comparison With Bare and Hec Coating Capillarysupporting

confidence: 84%

“…Therefore, in order to solve the insufficient adsorption of hydrophilic polymer on the capillary wall, a variety of positively charged polymers, such as polyethyleneimine [21,22]; a copolymer of vinylpyrrolidone and vinylimidazole [23]; Polybrene, poly(methoxyethoxyethyl) ethylenimine, poly(diallyldimethylammonium) chloride [24,25], ethylpyrrolidine methacrylate-N,N-dimethylacrylamide (EpyM-DMA) [26,27] were used as capillary coating for protein analysis by CE. Especially, cationic polysaccharides and their derivatives have attracted much attention for their both dynamic coating ability and reproducibility of separations [28][29][30][31][32][33]. In previous studies, the cationic polymer derivatives were shown to be capable of suppressing the adsorption of fluorescently labeled amino acids, peptides, and proteins.…”

Section: Introductionmentioning

confidence: 99%

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Cationized hydroxyethylcellulose as a novel, adsorbed coating for basic protein separation by capillary electrophoresis

et al. 2008

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We present cationized hydroxyethylcellulose (cat-HEC) synthesized in our laboratory as a novel physically adsorbed coating for CE. This capillary coating is simple and easy to obtain as it only requires flushing the capillary with polymer aqueous solution. A comparative study with and without polymers was performed. The adsorbed cat-HEC coating exhibited minimal interactions with basic proteins, providing efficient basic protein separations with excellent reproducibility. Under broad pHs, the amine groups are the main charged groups bringing about a global positive charge on the capillary wall. As a consequence, the cat-HEC coating produced an anodal EOF performance. A comparative study on the use of hydroxyethylcellulose (HEC) and cat-HEC as physically adsorbed coatings for CE are also presented. The separation efficiency and analysis reproducibility proved that the cat-HEC polymer was efficient in suppressing the adsorption of basic proteins onto the silica capillary wall. The long-term stability of the cat-HEC coating in consecutive protein separation runs has demonstrated the suitability of the coating for high-throughput electrophoretic protein separations.

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Section: Comparison With Bare and Hec Coating Capillarysupporting

confidence: 84%

Section: Introductionmentioning

confidence: 99%

Cationized hydroxyethylcellulose as a novel, adsorbed coating for basic protein separation by capillary electrophoresis

et al. 2008

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“…Recently, coatings with a copolymer (EpyM-DMA) synthesized from 2-ethyl-(2-pyrrolidine) methacrylate (EpyM) and N,N-dimethylacrylamide (DMA) [187] or with PolyE-323 [173,188], a novel polyamine, were both applied successfully in separating peptides and proteins in standard solutions and real samples. Contrary to PB, neither polymer bleeding or increase in MS background signal nor polymer degradation in the presence of MeOH or ACN was detected in consecutive CE-ESI-MS runs for the aforementioned new polymer generation [173,187].…”

Section: Capillary Coating In Cze-esi-msmentioning

confidence: 99%

Advances in the analysis of proteins and peptides by capillary electrophoresis with matrix‐assisted laser desorption/ionization and electrospray‐mass spectrometry detection

2005

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High throughput, outstanding certainty in peptide/protein identification, exceptional resolution, and quantitative information are essential pillars in proteome research. Capillary electrophoresis (CE) coupled to mass spectrometry (MS) has proven to meet these requirements. Soft ionization techniques, such as matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI), have paved the way for the story of success of CE-MS in the analysis of biomolecules and both approaches are subject of discussion in this article. Meanwhile, CE-MS is far away from representing a homogeneous field. Therefore the review will cover a vast area including the coupling of different modes of CE (capillary zone electrophoresis, capillary isoelectric foscusing, capillary electrochromatography, micellar electrokinetic chromatography, nonaqueous capillary electrophoresis) to MS as well as on-line preconcentration techniques (transient capillary isotachophoresis, solid-phase extraction, membrane preconcentration) applied to compensate for restricted detection sensitivity. Special attention is given to improvements in interfacing, namely addressing nanospray and coaxial sheath liquid design. Peptide mapping, collision-induced dissociation with subsequent tandem MS, and amendments in mass accuracy of instruments improve information validity gained from MS data. With 2-D on-line coupling of liquid chromatography (LC) and CE a further topic will be discussed. A special section is dedicated to recent attempts in establishing CE-ESI-MS in proteomics, in the clinical and diagnostic field, and in the food sector.

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“…The novel polymer, PolyE-323, recently reported by our research group showed good deactivation properties for the analysis of basic proteins [55]. Further, the polymer surface generated an anodal EOF of high magnitude, which makes exploration of PolyE-323-coated capillaries for use in CE-ESI-MS attractive.…”

Section: Resultsmentioning

confidence: 99%

A polyamine coating for enhanced capillary electrophoresis‐electrospray ionization‐mass spectrometry of proteins and peptides

et al. 2004

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A procedure for enhanced capillary electrophoresis-electrospray ionization-mass spectrometry (CE-ESI-MS) of proteins is presented. The use of a newly presented capillary coating, PolyE-323, provided fast separations of typically a few minutes with high efficiency, good deactivation, and no bleeding into the mass spectrometer. Capillaries coated with PolyE-323 showed high stability over a range of pH 2-10, and tolerance towards methanol and acetonitrile, two modifiers commonly used in CE-ESI-MS. Due to the speed and simplicity of the coating procedure, the polymeric surface could, if necessary, easily be regenerated. This capability is especially valuable when working with samples of complex matrix, where a capillary surface cleaning step might be desired in order to eliminate possible memory effects. The potential of PolyE-323-coated capillaries in bioanalysis using CE-ESI-MS was demonstrated by analyzing peptides and proteins up to 66 kDa using time of flight (TOF)-MS. Due to the stable, anodal electroosmotic flow generated by the coating, the use of a sheathless ESI interface was enabled, demonstrated in peptide analysis with attomole sensitivity. The fast on-line CE-ESI-TOF system using PolyE-323-coated capillaries provided efficient separation and detection of a large number of peaks in a short time, exemplified by the analysis of a tryptic digest of bovine serum albumin (BSA). The capability of the developed capillary surface coating was demonstrated by the separation of human plasma and cerebrospinal fluid (CSF).

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Novel polyamine coating providing non-covalent deactivation and reversed electroosmotic flow of fused-silica capillaries for capillary electrophoresis

Cited by 75 publications

References 31 publications

Cationized hydroxyethylcellulose as a novel, adsorbed coating for basic protein separation by capillary electrophoresis

Cationized hydroxyethylcellulose as a novel, adsorbed coating for basic protein separation by capillary electrophoresis

Advances in the analysis of proteins and peptides by capillary electrophoresis with matrix‐assisted laser desorption/ionization and electrospray‐mass spectrometry detection

A polyamine coating for enhanced capillary electrophoresis‐electrospray ionization‐mass spectrometry of proteins and peptides

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