High‐sensitive protein analysis by FESI‐CE‐MALDI‐MS

Pourhaghighi, Mohammad Reza; Busnel, Jean‐Marc; Girault, Hubert H.

doi:10.1002/elps.201100024

Cited by 22 publications

(21 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…CE‐UV/MALDI‐MS interface has been evaluated in terms of repeatability and robustness following the optimized separation conditions described by the team of Prof. Girault . As a first step, we have evaluated the external UV detection (Fig.…”

Section: Resultsmentioning

confidence: 99%

Analysis of monoclonal antibody by a novel CE‐UV/MALDI‐MS interface

et al. 2014

View full text Add to dashboard Cite

mAbs are highly complex proteins that present a wide range of microheterogeneity that requires multiple analytical methods for full structure assessment and quality control. As a consequence, the characterization of mAbs on different levels is particularly product- and time-consuming. CE-MS couplings, especially to MALDI, appear really attractive methods for the characterization of biological samples. In this work, we report the last instrumental development and performance of the first totally automated off-line CE-UV/MALDI-MS/MS. This interface is based on the removal of the original UV cell of the CE apparatus, modification of the spotting device geometry, and creation of an integrated delivery matrix system. The performance of the method was evaluated with separation of five intact proteins and a tryptic digest mixture of nine proteins. Intact protein application shows the acquisition of electropherograms with high resolution and high repeatability. In the peptide mapping approach, a total number of 154 unique identified peptides were characterized using MS/MS spectra corresponding to average sequence coverage of 64.1%. Comparison with NanoLC/MALDI-MS/MS showed complementarity at the peptide level with an increase of 42% when using CE/MALDI-MS coupling. Finally, this work represents the first analysis of intact mAb charge variants by CZE using an MS detection. Moreover, using a peptide mapping approach CE-UV/MALDI-MS/MS fragmentation allowed 100% sequence coverage of the light chain and 92% of the heavy chain, and the separation of four major glycosylated peptides and their structural characterization.

show abstract

Section: Resultsmentioning

confidence: 99%

Analysis of monoclonal antibody by a novel CE‐UV/MALDI‐MS interface

et al. 2014

View full text Add to dashboard Cite

show abstract

“…FASS-related techniques such as FASI and LVSS has also been applied to CE-MS [53][54][55][56][57][58][59][60][61].…”

Section: Field-amplified Sample Stacking and Related Techniquesmentioning

confidence: 99%

Recent Studies on Online Sample Preconcentration Methods inCapillary Electrophoresis Coupled with Mass Spectrometry

Kawai

2017

Chromatography

View full text Add to dashboard Cite

This paper reviews recent 5-years application researches using online sample preconcentration (OSP) methods in capillary electrophoresis (CE) coupled with mass spectrometry (MS). CE-MS has excellent analytical features such as high-resolution separation, rapid analysis time, low sample consumption, and small consumption of organic solvents. Recently, low flow-rate interface for electrospray ionization has been developed and zmol-level sensitivity has been achieved. However, application of CE-MS has still been prevented due to small capacity of sample injection volume. OSP methods have been developed to address this issue and up to 5,000-fold sensitivity improvement was obtained so far. Various application has been carried out such as proteomics, metabolomics, and glycomics, and CE-MS attracts much attention as the highest-performance analytical tool for small-volume bioanalysis. Taking account of further progress in CE-MS, smart and practical application of OSP methods will be more important in various analytical fields.

show abstract

“…In order to increase the sensitivity of CZE‐MALDI‐MS, Pourhaghighi et al developed a field‐enhanced sample injection strategy for intact proteins . Injections of intact proteins were performed electrokinetically for 8 min, which resulted in an increase of peak area between a factor 1000 and 8000 compared to normal electrokinetic injections.…”

Section: Ce‐maldi‐msmentioning

confidence: 99%

CE‐MS for the analysis of intact proteins 2010–2012

2012

View full text Add to dashboard Cite

Since its introduction in 1987, CE-MS has become an increasingly important technique for the analysis of biomolecules. Since our previous update on CE-MS methods within the field of intact protein analysis (Electrophoresis 2011, 32, 66-82), a variety of interesting methodological improvements and applications have been reported in literature. Therefore, this article presents an overview of the development and application of CE-MS for intact protein analysis as published between June 2010 and June 2012. The article is divided in sections that treat CE coupled to MS through ESI, MALDI, and ICP ionization, respectively. In the section about CE-ESI-MS, technological developments with respect to CE-MS interfacing, prevention of protein adsorption, and chip-based CE-MS are treated in more detail. Novel interfacing strategies and the development of improved capillary coating strategies appeared to be the major developments. Furthermore, in all sections, the applicability of CE-MS for intact protein analysis is demonstrated by representative examples, including important developments in the fields of biopharmaceutical characterization and the analysis of proteins in biological samples. Finally, some general conclusions and future perspectives are given.

show abstract

High‐sensitive protein analysis by FESI‐CE‐MALDI‐MS

Cited by 22 publications

References 38 publications

Analysis of monoclonal antibody by a novel CE‐UV/MALDI‐MS interface

Analysis of monoclonal antibody by a novel CE‐UV/MALDI‐MS interface

Recent Studies on Online Sample Preconcentration Methods inCapillary Electrophoresis Coupled with Mass Spectrometry

CE‐MS for the analysis of intact proteins 2010–2012

Contact Info

Product

Resources

About