A simple and inexpensive approach to interfacing high‐performance liquid chromatography and matrix‐assisted laser desorption/ionization‐time of flight‐mass spectrometry

Fung, Kim Y. C.; Ašković, Srdjan; Basile, Franco; Duncan, Mark W.

doi:10.1002/pmic.200300843

Cited by 25 publications

(23 citation statements)

References 31 publications

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“…The analysis of intact proteins is an important component of proteomic research as it yields information about post-translational modifications of proteins and the ability to detect other post-translational events leading to multiple products derived from a single gene 31 . Application of DESI-MS as an analysis tool for protein molecules in the field of proteomics is ideal due to its simplicity, speed of analysis for high throughput applications, and analytical specificity.…”

Section: Introductionmentioning

confidence: 99%

Desorption Electrospray Ionization-Mass Spectrometry of Proteins

et al. 2007

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Desorption electrospray ionization-mass spectrometry (DESI-MS) was evaluated for the detection of proteins ranging in molecular weight from 12 KDa to 66 KDa. Proteins were uniformly deposited on a solid surface without pretreatment and analyzed with a DESI source coupled to a quadrupole ion trap mass spectrometer. DESI parameters optimized for protein detection included solvent flow rate, temperature of heated capillary tube, incident and reflection angle, sheath gas pressure, and ESI voltage. Detection limits were obtained for all protein standards and they were found to decrease with decreasing protein molecular weight: for cytochrome-C (12.3 KDa) and lysozyme (14.3 KDa) a detection limit of 4 ng/mm 2 was obtained; for apomyoglobin (16.9 KDa) 20 ng/mm 2 ; for β-lactoglobulin B (18.2 KDa) 50 ng/mm 2 ; and for chymotrypsinogen-A (25.6 KDa) 100 ng/mm 2 . The DESI-MS analysis of higher molecular weight proteins such as ovalbumin (44.4 KDa) and bovine serum albumin (66.4 KDa) yielded mass spectra of low signal-to-noise ratio (S/N), making their detection and molecular weight determination difficult. In this study, DESI-MS proved to be a rapid and robust method for accurate MW determination for proteins up to 17 KDa under ambient conditions. Finally, we demonstrated the DESI-MS detection of the bacteriophage MS 2 capsid protein from crude samples with minimal sample preparation.

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Section: Introductionmentioning

confidence: 99%

Desorption Electrospray Ionization-Mass Spectrometry of Proteins

et al. 2007

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“…Although the use of tandem mass spectrometry can often provide confident identifications, there is increasing interest in higher throughput approaches that exploit highly accurate mass measurements [3][4][5][6][7][8]. Earlier studies have shown [9,10] that utilizing accurate mass spectrometric measurements for MS based identification of peptides within Ϯ0.1 ppm uncertainty (tolerance) can allow significant levels of confidence in protein identifications, even from mixtures with the complexity of some smaller eukaryotic systems (e.g., yeast).…”

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confidence: 99%

The Utility of Accurate Mass and LC Elution Time Information in the Analysis of Complex Proteomes

Norbeck

Monroe

Adkins

et al. 2005

J. Am. Soc. Mass Spectrom.

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The combination of mass and normalized elution time (NET) of a peptide identified by liquid chromatography-mass spectrometry (LC-MS) measurements can serve as a unique signature for that peptide. However, the specificity of an LC-MS measurement depends upon the complexity of the proteome (i.e., the number of possible peptides) and the accuracy of the LC-MS measurements. In this work, theoretical tryptic digests of all predicted proteins from the genomes of three organisms of varying complexity were evaluated for specificity. Accuracy of the LC-MS measurement of mass-NET pairs (on a 0 to 1.0 NET scale) was described by bivariate normal sampling distributions centered on the peptide signatures. Measurement accuracy (i.e., mass and NET standard deviations of Ϯ0.1, 1, 5, and 10 ppm, and Ϯ0.01 and 0.05, respectively) was varied to evaluate improvements in process quality. The spatially localized confidence score, a conditional probability of peptide uniqueness, formed the basis for the peptide identification. Application of this approach to organisms with comparatively small proteomes, such as Deinococcus radiodurans, shows that modest mass and elution time accuracies are generally adequate for confidently identifying most peptides. For more complex proteomes, more accurate measurements are required. However, the study suggests that the majority of proteins for even the human proteome should be identifiable with reasonable confidence by using LC-MS measurements with mass accuracies within Ϯ1 ppm and high efficiency separations having elution time measurements within Ϯ0.01

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“…The target deposition rate was 0.25 ml min Ϫ1 . A similar set up that has been developed and used previously for MALDI film deposition has been described elsewhere [9].…”

Section: Methodsmentioning

confidence: 99%

Clustering of pentacene and functionalized pentacene ions in a matrix-assisted laser desorption/ionization orthogonal TOF mass spectrometer

Shcherbyna

Böhme

Baranov

et al. 2006

J. Am. Soc. Mass Spectrom.

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A high-performance orthogonal time-of flight (TOF) mass spectrometer, in combination with the matrix assisted laser desorption/ionization (MALDI) source operating at elevated pressure ( approximately 1 torr in N(2)), was used to perform MALDI-TOF analyses of pentacene and some of its derivatives with and without an added matrix. These molecules are among the most interesting semiconductor materials for organic thin film transistor applications (OTFT). The observation of ion-molecule reactions between "cold" analyte ions and neutral analyte molecules in the gas phase has provided some insight into the mechanism of pentacene cluster formation and its functionalized derivatives. Furthermore, some of the matrices employed to assist the desorption/ionization process of these compounds were observed to influence the outcome via ion-molecule reactions of analyte ions and matrix molecules in the gas phase. The stability and reactivity of the compounds and their clusters in the MALDI plume during gas-phase expansion were evaluated; possible structures of the resulting clusters are discussed. The MALDI-TOF technique was also helpful in distinguishing between two isomeric forms of bis-[(triisopropylsilyl)-ethynyl]-pentacene.

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A simple and inexpensive approach to interfacing high‐performance liquid chromatography and matrix‐assisted laser desorption/ionization‐time of flight‐mass spectrometry

Cited by 25 publications

References 31 publications

Desorption Electrospray Ionization-Mass Spectrometry of Proteins

Desorption Electrospray Ionization-Mass Spectrometry of Proteins

The Utility of Accurate Mass and LC Elution Time Information in the Analysis of Complex Proteomes

Clustering of pentacene and functionalized pentacene ions in a matrix-assisted laser desorption/ionization orthogonal TOF mass spectrometer

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