Analysis of non-covalent protein complexes by capillary electrophoresis–time-of-flight mass spectrometry

Brenner‐Weiß, Gerald; Kirschhöfer, Frank; Kühl, Boris; Nusser, Michael; Obst, U.

doi:10.1016/s0021-9673(03)00553-3

Cited by 34 publications

(35 citation statements)

References 7 publications

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“…Selecting appropriate ES conditions, intact holo-Mb with the heme group still noncovalently attached can be detected in CZE-ESI-TOF-MS, although both a compact and a more relaxed holo-Mb conformation can be distinguished. Increasing the MeOH content in the SL shifts the equilibrium towards the apo-Mb form [142]. Ammonium acetate is known to shift CSD towards higher m/z values and the effect has been related to basicity of acetate, which favors removal of protons from proteins [72,139,141], but also to the loss of NH 4…”

Section: Relation Between Charge State Distribution and Protein Confomentioning

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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Section: Relation Between Charge State Distribution and Protein Confomentioning

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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“…CZE was used to study b-amyloid and its aggregation [275], botulinum neurotoxin B light chain activity [276], nonenzymatic posttranslation modifications on BSA [277], aminoacyl-tRNA synthetase localized in cell nucleus [278], protein-protein interactions [279], aggregation and denaturation of antibodies [280], glycosylation of murine erythropoietin [281], PEGylation of proteins [282], glutamine deamidation in proteins [283], complexation of apomyoglobin with Coumarin 153 [284], reaction of FQ with ovalbumin [285], interaction of GFP with calcium [286], noncovalent complexation in myoglobin [287], DNA-protein interactions [288][289][290], binding fucoidan to antithrombin [291], retinoic acid to b-lactoglobulin B [292], and binding phospholipid and glycosaminoglycan to human b 2 -glycoprotein [293].…”

Section: Other Proteinsmentioning

confidence: 99%

Capillary electrophoresis of proteins 2003–2005

Dolnı́k¹

2006

Electrophoresis

139

117

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This review article with 304 references describes recent developments in CE of proteins, and covers the two years since the previous review (Hutterer, K., Dolník, V., Electrophoresis 2003, 24, 3998-4012) through Spring 2005. It covers topics related to CE of proteins, including modeling of the electrophoretic migration of proteins, sample pretreatment, wall coatings, improving separation, various forms of detection, special electrophoretic techniques such as affinity CE, CIEF, and applications of CE to the analysis of proteins in real-world samples including human body fluids, food and agricultural samples, protein pharmaceuticals, and recombinant protein preparations.

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“…Corona formation was the central problem, stemming from an overpotential. The use of spray potentials as low as 0 V has been reported [20], however, even with a zero spray potential applied and the electrospray needle moved as far from the spray shield as possible, corona formation was still observed. It was concluded that due to this fact, TFA in the concentrations required for the separation of the selected resin samples was unsuitable as a BGE for this instrument.…”

Section: Optimization Of Parametersmentioning

confidence: 99%

Analysis of melamine resins by capillary zone electrophoresis with electrospray ionization‐mass spectrometric detection

2005

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A method for the determination of the major components of (methoxymethyl)melamine resins, with quantitative analysis of unreacted melamine by capillary zone electrophoresis (CZE) using electrospray ionization-mass spectrometry (ESI-MS) is presented. Using a low background electrolyte (BGE) pH, components are separated according to their charge/ionic radius ratio with a distinctly different separation selectivity compared to the HPLC methods commonly employed in melamine-resin analysis. The use of a time-of-flight mass spectrometer (TOF-MS) was concluded to be necessary, as the complex samples studied required maximum sensitivity and resolution, which is clearly superior for TOF-MS detectors over their quadrupole counterparts. A standard curve of free melamine was determined with an R(2) = 0.999 over a concentration range of an order of magnitude. This method offers the unique separation selectivity of CZE as well as a quicker analysis time, especially for dimers compared to the HPLC methods used to date.

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Analysis of non-covalent protein complexes by capillary electrophoresis–time-of-flight mass spectrometry

Cited by 34 publications

References 7 publications

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

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

Capillary electrophoresis of proteins 2003–2005

Analysis of melamine resins by capillary zone electrophoresis with electrospray ionization‐mass spectrometric detection

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