Advances in the analysis of proteins and peptides by capillary electrophoresis with matrix‐assisted laser desorption/ionization and electrospray‐mass spectrometry detection

Stutz, Hanno

doi:10.1002/elps.200410130

Cited by 161 publications

(175 citation statements)

References 276 publications

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“…The initial step is the same as proposed for the w 3 ϩ fragment formation in Scheme 1, i.e., the loss of the neutral guanine, induced through its initial protonation. Like in the mechanism shown in Scheme 1 the 1,2-elimination initiated by the base loss is completed by a proton transfer from the 2=-carbon of the sugar moiety to the next neighboring base with the highest proton affinity.…”

Section: -B 4 H]mentioning

confidence: 99%

“…Especially in the field of proteomics, these methods have proven to be very powerful both for the identification of known proteins and for the sequencing of unknown proteins and peptides. The metastable decay of preselected parent ions in MALDI has also proven to be very useful in proteomics [1][2][3], and is today a key technique in sequence analysis in proteomics. However, at the same time, the performance of these methods has not met expectations when it comes to the analyses of DNA segments.…”

mentioning

confidence: 99%

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Sodium controlled selective reactivity of protonated deoxy-oligonucleotides in the gas phase

Flosadóttir

Staňo

Ingólfsson

2008

J. Am. Soc. Mass Spectrom.

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Metastable fragmentation of the positively charged, hexameric oligonucleotides 5=-d(TTXYTT) (X and Y are dC, dG, or dA) and 5=-d(CTCGTT), 5=-d(TTCGTC) and 5=-d(CTCGTC) is studied after matrix assisted laser desorption/ionization (MALDI). The influence of the degree of sodiation, i.e., when the acidic protons are one by one exchanged against sodium ions, is systematically studied for the exchange of up to seven protons against sodium ions. Exchanging the acidic protons against sodium gradually quenches the backbone cleavage through the w and a-B channels, and quantitative quenching of these channels is generally achieved with the exchange of four protons against sodium ions. At the same time, the exchange of protons against sodium ions promotes the loss of a neutral, high proton affinity base. The formation of the w and a-B fragments is found to be highly dependent on the sequence of the central bases. A single mechanism consistent with these observations is proposed. In addition to the quenching of the classical w and a-B reaction channels, a drastic and abrupt on/off-switching of new reaction channels is observed as the degree of sodiation successively increases. These channels involve selective loss of the two central bases and the excision of a phosphodiester group and a sugar unit from the center of the oligonucleotides. Synchronously, the two terminal fragments recombine to form a tetramer containing the two terminal nucleosides from each end of the hexamer. Possible mechanism explaining these remarkable channels are discussed. T he need for new analytical methods and mass spectrometric techniques to shed light on the complex molecular structure and composition of biologically relevant molecules has drawn the attention of physicists and physical chemists more in the direction of gas-phase studies on molecules such as DNA, RNA, proteins, and peptides. The introduction of the electrospray ionization (ESI) and the matrix-assisted laser desorption ionization (MALDI) in the early 90s and the further development of these methods throughout the last decade have revolutionized the studies of biologically relevant molecules. Especially in the field of proteomics, these methods have proven to be very powerful both for the identification of known proteins and for the sequencing of unknown proteins and peptides. The metastable decay of preselected parent ions in MALDI has also proven to be very useful in proteomics [1][2][3], and is today a key technique in sequence analysis in proteomics. However, at the same time, the performance of these methods has not met expectations when it comes to the analyses of DNA segments. One of the difficulties hampering the use of MALDI TOF-MS in DNA analyses is the metastable and prompt decay of these molecules in the MALDI process [4 -11]. Furthermore, the strong affinity of oligonucleotides to exchange their numerous acidic protons against sodium complicates the sample preparation considerably and often sets limits to the achievable resolution [4,5]. In the last decades, several stud...

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Section: -B 4 H]mentioning

confidence: 99%

mentioning

confidence: 99%

Sodium controlled selective reactivity of protonated deoxy-oligonucleotides in the gas phase

Flosadóttir

Staňo

Ingólfsson

2008

J. Am. Soc. Mass Spectrom.

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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].…”

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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“…Recentemente, trabalhos envolvendo estudos proteômicos são encontrados na literatura, os quais comprovam a evolução de CE-MS aplicada neste tipo de estudos. [111][112][113][114][115] A técnica de CE-MS também pode ser empregada na análise de resíduos de pesticidas em alimentos. O limite de detecção obtido é pelo menos uma ordem de grandeza menor que em CE-UV e, concomitantemente, menor que os permitidos por lei (limite máximo permitido para o resíduo) aumentando, portanto, a aplicabilidade desta técnica.…”

Section: 87-90unclassified

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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Advances in the analysis of proteins and peptides by capillary electrophoresis with matrix‐assisted laser desorption/ionization and electrospray‐mass spectrometry detection

Cited by 161 publications

References 276 publications

Sodium controlled selective reactivity of protonated deoxy-oligonucleotides in the gas phase

Sodium controlled selective reactivity of protonated deoxy-oligonucleotides in the gas phase

Capillary electrophoresis of proteins 2003–2005

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

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