Capillary electrophoresis – mass spectrometry: 15 years of developments and applications

Schmitt‐Kopplin, Philippe; Frommberger, Moritz

doi:10.1002/elps.200305659

Cited by 282 publications

(237 citation statements)

References 386 publications

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“…The use of a hydroxypropylmethylcellulose-coated column significantly improved the separation of basic histones. A simple dialysis step prior to CE separation and the use of coaxial sheath liquid flow between the CE and ESI-MS interface made the online CE-ESI-MS possible [108][109][110][111]. Despite the potential power of the approach, CE-MS of histones is difficult due to:…”

Section: Hp Capillary Electrophoresis-msmentioning

confidence: 99%

Mass spectrometry-based strategies for characterization of histones and their post-translational modifications

Ren

Freitas

2007

Expert Review of Proteomics

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Due to the intimate interactions between histones and DNA, the characterization of histones has become the focus of great attention. A series of mass spectrometry-based technologies have been dedicated to the characterization and quantitation of different histone forms. This review focuses on the discussion of mass spectrometry-based strategies used for the characterization of histones and their post-translational modifications. Keywordshistone; mass spectrometry; post-translational modification Histones are a group of highly conserved proteins throughout eukaryotic evolution [1]. The core histones (H4, H3, H2A and H2B) form an octamer, in which a H2A-H2B dimer associates on each side of the H3-H4 tetramer through an interaction between the C-terminal halves of H2B and H4 [2]. The negatively charged DNA superhelix wraps around the histone octamer to form repeating nucleosome cores, which further assemble into higher order chromatin fibers stabilized by the linker histones (H1 and H5) and linker DNA [3]. The histone fold regions in core histones are highly conserved α-helices, which are connected by two loops. The third histone structural motif comprises the flexible and irregular histone tails, which are extended and reach outside of the nucleosomal unit to interact with DNA [4]. Challenge of molecular characterization: post-translational modifications & sequence variationsAs a building block of the nucleosome, histones play an important role in chromatin assembly and affect the interactions between DNA and other chromatin-associated proteins. Histones regulate gene transcription through their diverse post-translational modifications (PTMs), which include lysine acetylation, lysine and arginine methylation, serine and threonine phosphorylation, lysine ubiquitination, lysine sumoylation, and poly-ADP-ribosylation of glutamic acid [5][6][7][8]. Among all the PTMs, histone acetylation and methylation have been most †

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Section: Hp Capillary Electrophoresis-msmentioning

confidence: 99%

Mass spectrometry-based strategies for characterization of histones and their post-translational modifications

Ren

Freitas

2007

Expert Review of Proteomics

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“…Porém, a combinação de artefatos, tais como o uso de pressão na separação e aditivos no eletrólito, aliados a ferramentas computacionais possibilitou a análise eficiente e a quantificação de aminoácidos em diferentes matrizes. 2,69,84,112 A extração em fase sólida previamente à análise por CE-MS contribui para o aumento da detectabilidade dos métodos de análise de alimentos, como leite de soja, 118 pêssego, nectarina, 119 sucos de uva e laranja. 120 Um estudo envolvendo a avaliação de diferentes métodos de hidrólises da proteína albumina soro bovina (BSA -bovine serum albumin) usando CE-MS para análise dos aminoácidos resultantes mostrou a utilidade e aplicabilidade da técnica, empregando a castanha do pará como amostra real.…”

Section: 87-90unclassified

“…Porém a maior contribuição que a CE oferece é sua complementaridade aos métodos já estabelecidos, devido à variedade de modos de separação existentes -e muitos ainda em desenvolvimento -os quais permitem a análise de praticamente todas as classes de compostos. [1][2][3] A CE acoplada à espectrometria de massas (CE-MS) surgiu, aproximadamente, há duas décadas, e foi e vem sendo usada em diferentes aplicações, com crescente aceitação. O sucesso desta combinação alia o alto poder de resolução e eficiência da CE com a universalidade e sensibilidade que a MS oferece frente a outros sistemas de detecção como, por exemplo, espectrofotometria na região do UV-visível, bem como fluorescência induzida a laser (LIF -laser induced fluorescence), que são pouco informativos em relação à estrutura do analito.…”

Section: Introductionunclassified

Eletroforese capilar acoplada à espectrometria de massas (CE-MS): vinte anos de desenvolvimento

Assunção¹,

Bechara²,

Simionato³

et al. 2008

Quím. Nova

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Recebido em 29/1/08; aceito em 7/5/08; publicado na web em 8/10/08 CAPILLARY ELECTROPHORESIS COUPLED TO MASS SPECTROMETRY (CE-MS): TWENTY YEARS OF DEVELOPMENT. CE-MS has been increasingly used for analysis of a vast array of compounds. This article reviews the different electrophoretic modes, interfaces and mass analyzers that are commonly used in the CE-MS coupling, as well as the technique advantages and performance characteristics. A large compilation of CE-MS applications is also presented. Therefore, this review is both a guide for beginners and a collection of key references for people who are familiar to the technique. Furthermore, this is the first CE-MS review published in a Brazilian journal and marks the installation of the first two commercial CE-MS units in Sao Paulo State.Keywords: capillary electrophoresis; electrospray ionization; mass spectrometry. introdUÇÃoA eletroforese capilar (CE) vem ocupando um espaço crescente na química analítica como uma poderosa ferramenta de separação e identificação de uma ampla gama de moléculas. A CE possui alta resolução e eficiência, parâmetros estes de grande importância em um processo de separação. Outra vantagem é o baixo custo operacional: mesmo em condições em que se empregam solventes orgânicos, os volumes consumidos de amostra e eletrólito são desprezíveis comparados à cromatografia líquida de alta eficiência (HPLC), até mesmo quando a última é utilizada em escala capilar. Ressaltamos também que os processos eletroforéticos têm como característica positiva a pequena quantidade de amostra injetada (ordem de nL), fator de importância em aplicações clínicas. Porém a maior contribuição que a CE oferece é sua complementaridade aos métodos já estabelecidos, devido à variedade de modos de separação existentes -e muitos ainda em desenvolvimento -os quais permitem a análise de praticamente todas as classes de compostos.1-3 A CE acoplada à espectrometria de massas (CE-MS) surgiu, aproximadamente, há duas décadas, e foi e vem sendo usada em diferentes aplicações, com crescente aceitação. O sucesso desta combinação alia o alto poder de resolução e eficiência da CE com a universalidade e sensibilidade que a MS oferece frente a outros sistemas de detecção como, por exemplo, espectrofotometria na região do UV-visível, bem como fluorescência induzida a laser (LIF -laser induced fluorescence), que são pouco informativos em relação à estrutura do analito. Um exemplo de tais características de MS envolve seu uso juntamente a ferramentas computacionais, o que é altamente recomendado em estudos envolvendo amostras complexas (como proteínas, peptídeos e metabólitos), já que informações como massa molecular e estrutura (MS em tandem) são indispensáveis para caracterização e quantificação de tais analitos.Desta forma, as informações oferecidas pela segunda dimensão do detector de massas constituem uma outra vantagem do uso de CE-MS. Assim, solutos que apresentam pequenas variações no tempo de migração, ou até mesmo que migram com a mesma velocidade, podem ser identificados. Nest...

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“…Due to the advantages of CE over HPLC described in Section 1, efforts have been made to achieve stable CE-MS coupling [24,28,32]. The different methods of coupling and their technical problems were recently reviewed by Kolch et al [23] and are therefore not dealt with here in detail.…”

Section: Technical Aspects Of Ce-ms Couplingmentioning

confidence: 99%

Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposes

2005

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Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposesThe introduction of fast, sensitive, and robust techniques for proteomic analysis into clinical practice represents a major step toward a new diagnostic approach of body fluids. In addition, proteomics emerges as a key technology for the discovery of disease biomarkers in various body fluids. However, even in relatively protein-deprived body fluids such as urine, the complexity and wide dynamic range of protein expression pose a considerable challenge to both separation and identification technologies. In the present review we discuss from a clinical point-of-view recent advances of the use of proteomics in clinical diagnosis as well as therapy evaluation. We focus on capillary electrophoresis coupled to mass spectrometry (CE-MS) and discuss CE-MS from an application point of view evaluating its merits and vices with regard to biomarker discovery. This review further presents examples of clinical applications of CE-MS for detection and identification of biomarkers in urine. IntroductionIt is well appreciated for centuries that body fluids contain information on the "internal" environment, i.e., the function of tissues and organs, which can be used as a gauge for the well being of the organism. Although this concept was created by Hippocrates more than 2000 years ago, standardized clinical laboratory tests were not developed before the 20th century. Moreover, a systematic discovery of biomarkers was made possible only with the discovery of antibodies and their use in immunological assays such as radioimmuno assay (RIA), ELISA, etc. Their increasing use enabled detection and quantification of distinct molecules -mainly peptides and proteins -in various body fluids. However, these techniques were limited by their requirement of prior knowledge of the molecule and a specific probe. New functional proteomic approaches appeared to circumvent these limitations. Spurred by advances in MS it was hoped that proteomics would allow the analysis of thousands of peptides and proteins in one sample [1][2][3][4][5][6], but several obstacles turned up soon. The high complexity and wide dynamic range of peptides in body fluids required pre-MS separation techniques that offer high resolution and could cope with a wide dynamic range of peptide concentrations [3]. Further, suitable software for interpretation of a vast number of generated MS spectra as well as comparison of the data was a major challenge [7].The approach towards solving most of these problems was to reduce data complexity, for example by arbitrarily collecting data on only a few polypeptides and also to reduce resolution of the analysis instrument. Both were accomplished with the surface-enhanced laser desorption/ionization (SELDI) technology -a complex solution of analytes was allowed to interact with an active surface (ion-exchange, hydro-

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Capillary electrophoresis – mass spectrometry: 15 years of developments and applications

Cited by 282 publications

References 386 publications

Mass spectrometry-based strategies for characterization of histones and their post-translational modifications

Mass spectrometry-based strategies for characterization of histones and their post-translational modifications

Eletroforese capilar acoplada à espectrometria de massas (CE-MS): vinte anos de desenvolvimento

Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposes

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