Determination of bioactive peptides using capillary zone electrophoresis/mass spectrometry

Moseley, M. Arthur; Deterding, Leesa J.; Tomer, Kenneth B.; Jorgenson, James W.

doi:10.1021/ac00002a005

Cited by 148 publications

(71 citation statements)

References 53 publications

(54 reference statements)

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“…Columns were prepared as described by Moseley et al 17 Acid treatment was carried out with a 6 M solution of HCl for approximately 4 h and the column was purged with helium for 12 h at 300°C in a GC oven. Toluene was pumped through the column for 10 min, followed by a 2 h rinse was a 5% (v/v) solution of (3-aminopropyl)trimethoxysilane in toluene.…”

Section: Capillary Coatingsmentioning

confidence: 99%

Disposable Emitters for On-line Capillary Zone Electrophoresis/Nanoelectrospray Mass Spectrometry

Bateman

White

Thibault

1997

Rapid Commun. Mass Spectrom.

116

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A new method for preparing disposable microsprayers, for on-line capillary-zone electrophoresis/electrospray mass spectrometry (CZE/ES-MS) at sub-microliter per minute flow rates, is described. Different means of coupling these microsprayers to the CZE columns are presented, and compared with the more traditional one-piece capillary arrangement used for on-line CZE/ES-MS analysis. Enhancement of separation performance was obtained using capillaries coated with [(acryloylamino) propyl]trimethylammonium chloride reagents, an attractive alternative to other amine-coated capillaries. The use of disposable microsprayers provided ease of operation and flexibility for on-line CZE/ES-MS experiments, since CZE columns can be prepared independently from the microsprayers. The application of these microsprayers to unattended CZE/ES-MS operation for more than 8 hours was evaluated, and found to provide relative standard deviation values of 3-8% on peak areas and less than 1% on migration times. Full-scan mass spectral acquisition was obtained for low femtomol injections of peptides. The technique was applied to the analysis of a single chain lectin from Phaseolus vulgaris, using both CZE/ES-MS and CZE combined with tandem mass spectrometry. The mass spectral data obtained in this way provided information on the structure of glycopeptides identified in digests of the glycoprotein.

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Section: Capillary Coatingsmentioning

confidence: 99%

Disposable Emitters for On-line Capillary Zone Electrophoresis/Nanoelectrospray Mass Spectrometry

Bateman

White

Thibault

1997

Rapid Commun. Mass Spectrom.

116

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“…[20][21][22][23][24] We tested three different capillaries in this study. One of them was recovered with PVA, a highly hydrophilic polymer covalently bound to the capillary surface.…”

Section: Capillary Inner Coatingsmentioning

confidence: 99%

Evaluation of capillaries with different inner coatings for DNA analysis using dilute polymer solutions by capillary electrophoresis

Eugênio¹,

Carrilho²

2009

J. Braz. Chem. Soc.

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No presente trabalho, três colunas capilares, uma sem recobrimento interno e duas com recobrimentos internos diferentes foram utilizadas na separação de fragmentos de DNA: poli(vinil álcool) (PVA) e poli(dimetilacrilamida) (PDMA) -ambos de recobrimento covalente -foram comparados para a separação de DNA utilizando soluções poliméricas. As separações foram realizadas usando hidroxietilcelulose (HEC) (90-105 kDa) nas concentrações entre 0,00 e 2,00% m/v. Os resultados indicaram que a eficiência de separação foi maior no capilar de PVA do que no de PDMA, em todas as concentrações de HEC testadas. Ainda, uma resolução superior também foi observada com o capilar de PVA, já que com o capilar de PDMA o formato dos picos não se mostrou reprodutível quando corridas subseqüentes foram realizadas. Contrariamente ao relatado na literatura, nenhuma separação foi conseguida com o capilar sem revestimento interno.In this work three capillary columns, one with uncoated inner wall and two with covalentlybound internal coatings -poly(vinyl alcohol) (PVA) and poly(dimethylacrylamide) (PDMA) -both covalently covered -were used to separate DNA fragments and compared to DNA separation using replaceable polymer solutions. The separations were performed using hydroxyethylcellulose (HEC) (90-105 kDa) in concentrations ranging from 0.00 to 2.00% m/v. The results indicated that the separation efficiency was higher in the PVA capillary than in the PDMA in all evaluated concentrations of HEC. In addition, higher resolution was also observed in PVA-coated capillary since in PDMA the shape of the peaks was not reproducible when subsequent runs were performed. Contrary to what has previously been reported in the literature, no reasonable separation was possible in bare fused silica.Keywords: DNA separation, capillary coating, laser-induced fluorescence, PDMA, PVA IntroductionCapillary electrophoresis (CE) is a separation technique widely used for separation of biological macromolecules. It offers great advantages over slab-gel electrophoresis (SGE) -shorter analysis time, smaller quantity of sample, greater efficiency of separation, and easier automation. 1,2 The first attempts to apply CE to DNA separations were carried out in capillaries filled with cross-linked gels, like polyacrylamide. [3][4][5] Later on, some polymeric solutions were used, known as dynamic or physical gels, which are easily prepared and introduced into the capillary, thus enabling its renewal after each analysis and the use of a single capillary for many separations, without any problem with the subsequent separations. 6 DNA migration mechanisms in polymeric matricesThe Ogston and the reptation models were first proposed to explain DNA migration in slab gels, but they have also shown themselves suitable for polymeric solutions. 7 According to the Ogston theory, the DNA fragment keeps a spherical and rigid conformation of radius Rg. This particle moves across a random net formed by the gel fibers. These fibers form pores with average size ξ, which is a function of the po...

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“…Separation of bioactive peptides and subsequent analysis by fast atom bombardment-mass spectrometry (FAB-MS) utilize this silation to separate acidic proteins, but again a basic pH produces serious tailing for some proteins (38 have been used in several other laboratories with success (27,51). With the addition of PVA to the buffer, the walls became coated with this polymer and were deactivated or at least covered the reactive negative charge of the capillary wall.…”

Section: Coated Capillariesmentioning

confidence: 99%

Optimization of separation and detection schemes for DNA with pulsed field slab gel and capillary electrophoresis

McGregor

1993

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This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer.The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction.In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion.Oversize materials (e.g., maps, drawings, charts) are reproduced by sectioning the original, beginning at the upper left-hand corner and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book.

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Determination of bioactive peptides using capillary zone electrophoresis/mass spectrometry

Cited by 148 publications

References 53 publications

Disposable Emitters for On-line Capillary Zone Electrophoresis/Nanoelectrospray Mass Spectrometry

Disposable Emitters for On-line Capillary Zone Electrophoresis/Nanoelectrospray Mass Spectrometry

Evaluation of capillaries with different inner coatings for DNA analysis using dilute polymer solutions by capillary electrophoresis

Optimization of separation and detection schemes for DNA with pulsed field slab gel and capillary electrophoresis

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