Separations Combined with Mass Spectrometry

Tomer, Kenneth B.

doi:10.1021/cr990091m

Cited by 115 publications

(87 citation statements)

References 324 publications

(530 reference statements)

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“…These results demonstrate the utility of the AMT tag approach using capillary LC-TOF MS instruments, and also show that AMT tags developed using other instrumentation can be effectively utilized. T he two dominant separation methods used in proteomics are 2-dimensional gel electrophoresis [1] and, increasingly, liquid chromatography (LC) [2]. The first use of LC to obtain sequence information from proteins was by Fredrick Sanger in the 1940s and LC separation methods have experienced rapidly growing use since online LC-MS analysis based upon electrospray ionization (ESI) was demonstrated in the early 1990s [3][4][5][6][7].…”

mentioning

confidence: 99%

Proteome analyses using accurate mass and elution time peptide tags with capillary LC time-of-flight mass spectrometry

Strittmatter

Ferguson

Tang

et al. 2003

J. Am. Soc. Mass Spectrom.

148

113

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We describe the application of capillary liquid chromatography (LC) time-of-flight (TOF) mass spectrometric instrumentation for the rapid characterization of microbial proteomes. Previously (Lipton et al., Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 11049) the peptides from a series of growth conditions of Deinococcus radiodurans have been characterized using capillary LC MS/MS and accurate mass measurements which are captured as an accurate mass and time (AMT) tag database. Using this AMT tag database, detected peptides can be assigned using measurements obtained on a TOF due to the additional use of elution time data as a constraint. When peptide matches are obtained using AMT tags (i.e., using both constraints) unique matches of a mass spectral peak occurs 88% of the time. Not only are AMT tag matches unique in most cases, the coverage of the proteome is high; ϳ3500 unique peptide AMT tags are found on average per capillary LC run. From the results of the AMT tag database search, ϳ900 ORFs detected using LC-TOFMS, with ϳ500 ORFs covered by at least two AMT tags. These results indicate that AMT database searches with modest mass and elution time criteria can provide proteomic information for approximately one thousand proteins in a single run of Ͻ3 h. The advantage of this method over using MS/MS based techniques is the large number of identifications that occur in a single experiment as well as the basis for improved quantitation. For MS/MS experiments, the number of peptide identifications is severely restricted because of the time required to dissociate the peptides individually. These results demonstrate the utility of the AMT tag approach using capillary LC-TOF MS instruments, and also show that AMT tags developed using other instrumentation can be effectively utilized. (J Am Soc Mass Spectrom 2003, 14, 980 -991)

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confidence: 99%

Proteome analyses using accurate mass and elution time peptide tags with capillary LC time-of-flight mass spectrometry

Strittmatter

Ferguson

Tang

et al. 2003

J. Am. Soc. Mass Spectrom.

148

113

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“…However, three major difficulties have limited the coupling of LC to MS for many years, namely I) apparent incompatibility of the flow-rate as expressed by the need for introducing a large volume of a liquid effluent from a conventional LC column into the high vacuum of the mass spectrometer; II) incompatibility of the effluent composition as a result of the frequent use of non-volatile mobile phase additives during the LC separation process, and (III) ionization of non-volatile and/or thermally labile analytes (Niessen & Tinke, 1995). These difficulties were overcome with the advent of the atmospheric pressure ionization (API) techniques, more specifically electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) (Tomer, 2001). In ESI, the sample is dissolved in a polar, volatile solvent and pumped through a narrow stainless steel capillary (75-150 m i. d.).…”

Section: Why Use Lc-ms?mentioning

confidence: 99%

Electrospray Ionization Tandem Mass Spectrometry as a Tool for the Structural Elucidation and Dereplication of Natural Products: An Overview

Dias¹,

Melo²,

Crotti³

2012

Tandem Mass Spectrometry - Applications and Principles

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“…Mass spectrometry lacks intrinsic chemical selectivity, except for formula-specific identification by sufficiently high mass resolution (63,64), and differing drift velocities in ion mobility spectrometry. However, instruments in all of the areas named above have been interfaced to liquid chromatography, GC, and CE separations (67).…”

Section: Mass Spectrometrymentioning

confidence: 99%