Resolution power of two‐dimensional electrophoresis and identification of proteins from gels

Jungblut, Peter R.; Thiede, Bernd; Zimny‐Arndt, Ursula; Müller, Eva‐Christina; Scheler, Christian; Wittmann‐Liebold, Brigitte; Otto, Albrecht

doi:10.1002/elps.1150170505

Cited by 154 publications

(98 citation statements)

References 73 publications

(19 reference statements)

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“…The best separation is achieved by high-resolution 2-DE. Up to now, no other techniques have the high resolution power of this method [56]. It can be easily imagined, that a high resolution of complex protein mixtures is an essential step towards meaningful results in proteomics.…”

Section: Msmentioning

confidence: 99%

Two‐dimensional gel electrophoresis as tool for proteomics studies in combination with protein identification by mass spectrometry

2006

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The proteome analysis by 2-DE is one of the most potent methods of analyzing the complete proteome of cells, cell lines, organs and tissues in proteomics studies. It allows a fast overview of changes in cell processes by analysis of the entire protein extracts in any biological and medical research projects. New instrumentation and advanced technologies provide proteomics studies in a wide variety of biological and biomedical questions. Proteomics work is being applied to study antibiotics-resistant strains and human tissues of various brain, lung, and heart diseases. It cumulated in the identification of antigens for the design of new vaccines. These advances in proteomics have been possible through the development of advanced high-resolution 2-DE systems allowing resolution of up to 10 000 protein spots of entire cell lysates in combination with protein identification by new highly sensitive mass spectrometric techniques. The present technological achievements are suited for a high throughput screening of different cell situations. Proteomics may be used to investigate the health effects of radiation and electromagnetic field to clarify possible dangerous alterations in human beings.

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Section: Msmentioning

confidence: 99%

Two‐dimensional gel electrophoresis as tool for proteomics studies in combination with protein identification by mass spectrometry

2006

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“…Protein top-down analysis by mass spectrometry (MS) has provided efficient characterization of proteins in their intact forms including any "protein species" [1][2][3][4]. According to Schlüter and Jungblut et al [3], protein species are defined as the different proteins derived from a single gene.…”

Section: Introductionmentioning

confidence: 99%

“…sequence chain length and/or PTM) and includes recombinant proteins such as mAbs produced as drugs [1][2][3]. As such, we employed oligosaccharide profiling analysis for comprehensive glycan identification of an IgG1 mAb and a glycosylated IgG1 fusion protein.…”

Section: Introductionmentioning

confidence: 99%

Comprehensive glycosylation profiling of IgG and IgG-fusion proteins by top-down MS with multiple fragmentation techniques

Tran

Barton²,

Feng³

et al. 2016

Journal of Proteomics

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We employed top-and middle-down analyses with multiple fragmentation techniques including electron transfer dissociation (ETD), electron capture dissociation (ECD), and matrix-assisted laser desorption ionization in-source decay (MALDI-ISD) for characterization of a reference monoclonal antibody (mAb) IgG1 and a fusion IgG protein. Fourier transform ion cyclotron resonance (FT-ICR) or high performance liquid chromatography electrospray ionization (HPLC-ESI) on an Orbitrap was employed. These experiments provided a comprehensive view on the protein species; especially for different glycosylation level in these two proteins, which showed good agreement with oligosaccharide profiling. Top-and middle-down MS provided additional information regarding glycosylation sites and different combinational protein species that were not available from oligosaccharide mapping or conventional bottom-up analysis. Finally, incorporating a limited enzymatic digestion by immunoglobulin G-degrading enzyme of Streptococcus pyogene (IdeS) with MALDI-ISD analysis enabled extended sequence coverage of the internal region of protein without pre-fractionation. Biological significance: Oligosaccharide profiling together with top-and middle-down methods enabled: 1) detection of heterogeneous glycosylated protein species and sites in intact IgG1 and fusion proteins with high mass accuracy, 2) estimation of relative abundance levels of protein species in the sample, 3) confirmation of the protein termini structural information, and 4) improved sequence coverage by MALDI-ISD analysis for the internal regions of the proteins without sample pre-fractionation.

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“…Now the study of PTMs via MS-based proteomic strategies constitutes a vibrant and large field of research. Importantly, Jungblut and Schlüter proposed a systematic nomenclature for the comprehensive description of protein species and suggested that protein species sequence contain PTMs should replace the basic unit protein sequence [19][20][21]. Therefore, systematic investigation of PTMs could contribute to the holistic and comprehensive description of protein species and to elucidate potential biological roles of each protein species in cyanobacteria.…”

Section: Introductionmentioning

confidence: 99%

Proteomic analysis of post translational modifications in cyanobacteria

Xiong

Chen

2016

Journal of Proteomics

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Resolution power of two‐dimensional electrophoresis and identification of proteins from gels

Cited by 154 publications

References 73 publications

Two‐dimensional gel electrophoresis as tool for proteomics studies in combination with protein identification by mass spectrometry

Two‐dimensional gel electrophoresis as tool for proteomics studies in combination with protein identification by mass spectrometry

Comprehensive glycosylation profiling of IgG and IgG-fusion proteins by top-down MS with multiple fragmentation techniques

Proteomic analysis of post translational modifications in cyanobacteria

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