scispace - formally typeset
Journal ArticleDOI

Quantitative proteomic analysis by accurate mass retention time pairs.

TLDR
The principal focus of this paper will demonstrate the quantitative aspects of the methodology and continue with a discussion of the associated, complementary qualitative capabilities.
Abstract
Current methodologies for protein quantitation include 2-dimensional gel electrophoresis techniques, metabolic labeling, and stable isotope labeling methods to name only a few. The current literature illustrates both pros and cons for each of the previously mentioned methodologies. Keeping with the teachings of William of Ockham, “with all things being equal the simplest solution tends to be correct”, a simple LC/MS based methodology is presented that allows relative changes in abundance of proteins in highly complex mixtures to be determined. Utilizing a reproducible chromatographic separations system along with the high mass resolution and mass accuracy of an orthogonal time-of-flight mass spectrometer, the quantitative comparison of tens of thousands of ions emanating from identically prepared control and experimental samples can be made. Using this configuration, we can determine the change in relative abundance of a small number of ions between the two conditions solely by accurate mass and retention...

read more

Citations
More filters
Journal ArticleDOI

Andromeda: a peptide search engine integrated into the MaxQuant environment

TL;DR: A novel peptide search engine using a probabilistic scoring model that can handle data with arbitrarily high fragment mass accuracy, is able to assign and score complex patterns of post-translational modifications, and accommodates extremely large databases.
Journal ArticleDOI

Quantitative mass spectrometry in proteomics: a critical review

TL;DR: This review critically examine the more commonly used quantitative mass spectrometry methods for their individual merits and discusses challenges in arriving at meaningful interpretations of quantitative proteomic data.
Journal ArticleDOI

Absolute Quantification of Proteins by LCMSE A Virtue of Parallel ms Acquisition

TL;DR: A new method of absolute quantification of proteins is described, based on the discovery of an unexpected relationship between MS signal response and protein concentration, which is constant within a coefficient of variation of less than ±10%.
Journal ArticleDOI

Protein Analysis by Shotgun/Bottom-up Proteomics

TL;DR: The progress of proteomics has been driven by the development of new technologies for peptide/protein separation, mass spectrometry analysis, isotope labeling for quantification, and bioinformatics data analysis.
Journal ArticleDOI

Orthogonality of separation in two-dimensional liquid chromatography.

TL;DR: The RP-RP system (employing significantly different pH in both RP separation dimensions) had the highest practical peak capacity of 2D-LC systems investigated and was found to provide suitable orthogonality.
References
More filters
Journal ArticleDOI

Maximum entropy image reconstruction: general algorithm

TL;DR: Le maximum d'entropie (LDE) as mentioned in this paper is a technique optimale de reconstruction d'image, largement applicable en astronomie, and it is capable of generer des images provenant d'une large variete de types de donnees.
Journal ArticleDOI

Matrix effect in quantitative LC/MS/MS analyses of biological fluids: a method for determination of finasteride in human plasma at picogram per milliliter concentrations.

TL;DR: Development of a highly sensitive assay for I in human plasma at low picogram per milliliter concentrations using LC/MS/MS with a heated nebulizer (HN) interface, instead of a TISP interface, is described, practically eliminating the ion suppression.
Journal ArticleDOI

Metabolic labeling of C. elegans and D. melanogaster for quantitative proteomics

TL;DR: In this article, the authors describe quantitative 15N metabolic labeling of the multicellular organisms Caenorhabditis elegans, a nematode and Drosophila melanogaster, the common fruit fly, achieved by feeding them on 15N-labeled Escherichia coli and yeast, respectively.
Journal ArticleDOI

18O labeling: a tool for proteomics.

TL;DR: The evaluation of the relative Ziptip efficiency indicated a loss in sample recovery as the peptide concentration was reduced using normal conditions, suggesting that there is a limit below which there are diminishing returns.
Journal ArticleDOI

Ion suppression effects in liquid chromatography-electrospray-ionisation transport-region collision induced dissociation mass spectrometry with different serum extraction methods for systematic toxicological analysis with mass spectra libraries.

TL;DR: It could be demonstrated, that ion suppression is not generally present at any retention time when using reversed-phase HPLC with rather long gradient programs, but may play an important role in case of high-throughput LC-MS analysis, when the analyte is not separated from the LC-front, or in flow injection analysis without chromatographic separation.
Related Papers (5)