On‐line capillary electrophoresis‐mass spectrometry for the analysis of biomolecules

Hernández-Borges, Javier; Neusüß, Christian; Cifuentes, Alejandro; Pelzing, Matthias

doi:10.1002/elps.200405954

Cited by 189 publications

(153 citation statements)

References 208 publications

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“…1 in [33]) This form of ionization is essentially comparable with the micro-or nano-flow ionspray (depending on the liquid flow rate). The sample is diluted by the use of sheath liquid; but to a lesser extent than expected, probably due to incomplete mixing in the Taylor cone [24]. The sensitivity is generally superior with sheathless interfaces, but these appear to be less stable, at least in part due to deposits on the metal or graphite coating used.…”

Section: Technical Aspects Of Ce-ms Couplingmentioning

confidence: 95%

“…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%

“…Currently, the highest resolution in peptide separation can be accomplished by either combining 2-D LC-based technologies, or in a single step using CE. While HPLC is more widely used (for details see [1,20,21]), CE appears to offer several advantages: (i) it provides stable constant flow avoiding a gradient, which could result in changes in the ideal ionization parameters [22], (ii) it is fairly robust and uses inexpensive capillaries [23], (iii) it is compatible with essentially all buffers and analytes [24], and (iv) it provides fast separation and high resolution [25].…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Technical Aspects Of Ce-ms Couplingmentioning

confidence: 95%

Section: Technical Aspects Of Ce-ms Couplingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposes

2005

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“…Soft ionization techniques such as MALDI and ESI allowed a combination of CE and MS [99][100][101][102][103]. In CE-MS the proteins separated by CE are typically introduced into MS by an electrospray [104]. Recently, some off-line combinations with MALDI have been introduced [102].…”

Section: Msmentioning

confidence: 99%

Capillary electrophoresis of proteins 2003–2005

Dolnı́k¹

2006

Electrophoresis

139

117

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This review article with 304 references describes recent developments in CE of proteins, and covers the two years since the previous review (Hutterer, K., Dolník, V., Electrophoresis 2003, 24, 3998-4012) through Spring 2005. It covers topics related to CE of proteins, including modeling of the electrophoretic migration of proteins, sample pretreatment, wall coatings, improving separation, various forms of detection, special electrophoretic techniques such as affinity CE, CIEF, and applications of CE to the analysis of proteins in real-world samples including human body fluids, food and agricultural samples, protein pharmaceuticals, and recombinant protein preparations.

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“…Neste cenário, a CE-MS vem emergindo como uma ferramenta de grande impacto em bioanalítica, porém ainda são necessários grandes investimentos em pesquisa e desenvolvimento para atingir sua plena capacidade tecnológica. [4][5][6][7] instrUmEntaÇÃo Em 1987, Olivares et al relataram o uso inédito do espectrô-metro de massas como detector para CE. 8 Desde então, a técnica CE-MS tem recebido crescente aceitação, onde o MS complementa, ou substitui, detectores convencionais que são menos informativos e universais.…”

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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On‐line capillary electrophoresis‐mass spectrometry for the analysis of biomolecules

Cited by 189 publications

References 208 publications

Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposes

Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposes

Capillary electrophoresis of proteins 2003–2005

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

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