2015
DOI: 10.1002/elps.201500185
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Capillary isoelectric focusing‐mass spectrometry: Coupling strategies and applications

Abstract: IEF on immobilized pH gradient strips is a widespread tool for protein separation, especially as first dimension in commonly utilized 2DE. In the latter arrangement, separations are based on two orthogonal molecular characteristics according to pI in the first and molecular weight in the second dimension. However, the approach is time consuming, quantification is difficult and MS can be applied only offline. Capillary IEF and related IEF techniques in combination with MS provide similar information. The major … Show more

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Cited by 53 publications
(56 citation statements)
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“…CIEF online coupled to MS requires volatile electrode solutions, such as formic acid and ammonium hydroxide . In this simulation work and before , H 3 PO 4 and NaOH were used as anolyte and catholyte, respectively.…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…CIEF online coupled to MS requires volatile electrode solutions, such as formic acid and ammonium hydroxide . In this simulation work and before , H 3 PO 4 and NaOH were used as anolyte and catholyte, respectively.…”
Section: Resultsmentioning
confidence: 99%
“…Carrier ampholytes are synthetic mixtures of low molecular mass amphoteric components that possess high buffering capacity, solubility at the p I , good conductivity at the p I , and no biological effects . High efficiency, high selectivity, short analysis time, and low consumption of reagents are some of the recognized characteristics of this method performed in a capillary . In carrier ampholyte based CIEF sample components are typically applied as a zone mixed with the carrier ampholytes.…”
Section: Introductionmentioning
confidence: 99%
“…Coupling strategies with MS and applications of capillary IEF were reviewed by Hühner et al. . They emphasized important coupling techniques, which allow sensitive MS detection as well as proteomic and biopharmaceutical applications.…”
Section: Development Of Methodology and Instrumentationmentioning
confidence: 99%
“…The use of MS as the detection system provides structural information about the detected analytes allowing accurate relative molecular mass determination for proteins and polypeptides. Offline and online hyphenation of CIEF with MS have been reviewed recently . Several approaches were developed for analyses by MALDI‐TOF–MS after CIEF using, e.g., a sheath flow tee interface for the sample deposition to the MALDI plate , however, the online coupling provided more opportunities for the detection.…”
Section: Introductionmentioning
confidence: 99%
“…Offline and online hyphenation of CIEF with MS have been reviewed recently . Several approaches were developed for analyses by MALDI‐TOF–MS after CIEF using, e.g., a sheath flow tee interface for the sample deposition to the MALDI plate , however, the online coupling provided more opportunities for the detection. Clark and Naylor compared the separation of standard proteins analyzed by UV or MS detection after IEF and demonstrated the possibility of absolute identification for physiological proteins within complex in vivo derived mixtures .…”
Section: Introductionmentioning
confidence: 99%