A simple and robust conductive graphite coating for sheathless electrospray emitters used in capillary electrophoresis/mass spectrometry

Nilsson, Stefan; Wetterhall, Magnus; Bergquist, Jonas; Nyholm, Leif; Markides, Karin E.

doi:10.1002/rcm.466.abs

Cited by 35 publications

(63 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A total mobile phase flow of 250 µL/min (standard peptides) or 1 mL/min (BSA digest) from the two pumps was split approximately 1:1000 (standard peptides) or 1:250 (BSA digest) prior to the column. The eluent passed a UV detector prior to ESI in a novel sheathless electrospray emitter17 connected to an Analytica atmosphere‐vacuum interface (Analytica, Branford, CT, USA). The peptides were separated by a 28‐min gradient from 20.0% to 99.5% CH 3 CN (standard peptides) or 20.0% to 95.0% (BSA digest) CH 3 CN in H 2 O, 0.5% HAc.…”

Section: Methodsmentioning

confidence: 99%

Liquid chromatography and electron‐capture dissociation in Fourier transform ion cyclotron resonance mass spectrometry

Palmblad

Tsybin

Ramström

et al. 2002

Rapid Comm Mass Spectrometry

Self Cite

View full text Add to dashboard Cite

Liquid separation methods in combination with electrospray mass spectrometry as well as the recently introduced fragmentation method electron capture dissociation (ECD) have become powerful tools in proteomics research. This paper presents the results of the first successful attempts to combine liquid chromatography (LC) and Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS) with ECD in the analysis of a mixture of standard peptides and of a bovine serum albumin tryptic digest. A novel electron injection system provided conditions for ECD sufficient to yield extensive sequence information for the most abundant peptides in the mixtures on the time-scale of the chromatographic separation. The results suggest that LC/ECD-FTICRMS can be employed in the characterization of peptides in enzymatic digests of proteins or protein mixtures and identify and localize posttranslational modifications.

show abstract

Section: Methodsmentioning

confidence: 99%

Liquid chromatography and electron‐capture dissociation in Fourier transform ion cyclotron resonance mass spectrometry

Palmblad

Tsybin

Ramström

et al. 2002

Rapid Comm Mass Spectrometry

Self Cite

View full text Add to dashboard Cite

show abstract

“…Two HPLC pumps (JASCO 1580, JASCO Japan) delivered mobile phase gradients using solvent A (ACN:H 2 O:HAc -5:94.5:0.5), and solvent B (ACN:H 2 O:HAc -94.5:5:0.5). The peptides were electrosprayed on-line to a 9.4 T Fourier transform ion cyclotron resonance mass spectrometer (FTICR-MS; Bruker Daltonics, Billerica MA, USA) using a Black Dust (polyimide graphite) sheathless electrospray emitter [27] at a flow rate of 1 µL/min. The experimental setup has been described in detail elsewhere [28].…”

Section: In-gel Tryptic Digestion and Msmentioning

confidence: 99%

Targeted protein pullout from human tissue samples using competitive elution

Falk

Ramström

Eriksson

et al. 2010

Biotechnology Journal

View full text Add to dashboard Cite

One commonly used strategy to gain information on the proteins in a cell is to isolate the proteins of interest by specific binders, often antibodies. Not only the specificity of the capturing antibodies but also the washing and elution conditions are crucial to avoid false-positive protein identifications. Eluting the target protein from the matrix, while avoiding the release of unrelated background proteins, should both provide more correct information on the target protein and its interaction partners, and minimize the effort to perform downstream analyses through the reduced number of eluted proteins. In this study, a novel approach for selective protein pullout is presented. Monospecific antibodies were used to selectively pullout target proteins from a complex biosample. Subsequently, the target proteins were competitively eluted from the affinity media with the recombinant antigen. To deplete the antigen from the eluted sample, IMAC spin columns were utilized to bind the N-terminal His-tag of the antigens. The competitive elution method was applied both to a model system, and for the extraction of a native human target protein. In the model system the recombinant target protein BBC7 was spiked into a protein extract of human liver, whereas an endogenously expressed target protein, cTAGE5, was extracted from the liver extract directly. SDS-PAGE analysis and mass spectrometry confirmed affinity isolation of expected target proteins. More selective elution was obtained using the competitive procedure as compared to elution at low pH. Competitive elution has thus been shown to offer an effective approach for wide-scale pullout experiments where proteins and their interaction partners are to be studied.

show abstract

“…The conductive outer surface was obtained by coating conductive material such as gold, silver, copper, nickel or graphite materials. [6][7][8][9][10][11][12][13][14][15][16][17] Despite the improved sensitivity afforded by the sheathless design, the robustness was still challenge because of the poor stability of the metal coating and the electrolytic reaction within the tip.…”

Section: Introductionmentioning

confidence: 99%

Study of the electrical connection mechanism of sheathless interface for capillary electrophoresis‐electrospray ionization‐mass spectrometry

Tie

Zhang

Chen³

et al. 2012

J. Mass. Spectrom.

View full text Add to dashboard Cite

With the combination of high separation ability of capillary electrophoresis (CE) and strong identification ability of mass spectrometry (MS), CE/MS is becoming a powerful tool for polar and ionic analytes analysis. Different interfaces have been developed to enhance the sensitivity and reliability since the first introduction of CE/MS in 1987. A sheathless porous interface based on a new ions transferring electric connection technique was reported to be with high sensitivity and reliability. In this work, a series of optical and electrochemical experiments were designed to study the electric connection process. The results indicated that closing CE electrical circuit and applying MS spray voltage were achieved by the small ions transferring through the interface porous wall. The new electric connection method significantly enhanced the sensitivity, resolution and stability of the CE/MS analysis. The interface was applied in CE/MS detection of morphine and 6-monoacetylmorphine in urine sample and showed an equal sensitivity to LC/MS. With the significant improvement of sensitivity and stability, the CE/MS with the new interface showed strong potential for the determination of low abundance analytes.

show abstract

A simple and robust conductive graphite coating for sheathless electrospray emitters used in capillary electrophoresis/mass spectrometry

Cited by 35 publications

References 0 publications

Liquid chromatography and electron‐capture dissociation in Fourier transform ion cyclotron resonance mass spectrometry

Liquid chromatography and electron‐capture dissociation in Fourier transform ion cyclotron resonance mass spectrometry

Targeted protein pullout from human tissue samples using competitive elution

Study of the electrical connection mechanism of sheathless interface for capillary electrophoresis‐electrospray ionization‐mass spectrometry

Contact Info

Product

Resources

About