| Literature DB >> 26867742 |
Ioanna Ntai1,2,3, Timothy K Toby1,2,3, Richard D LeDuc1,2,3, Neil L Kelleher4,5,6.
Abstract
Biomarker discovery in the translational research has heavily relied on labeled and label-free quantitative bottom-up proteomics. Here, we describe a new approach to biomarker studies that utilizes high-throughput top-down proteomics and is the first to offer whole protein characterization and relative quantitation within the same experiment. Using yeast as a model, we report procedures for a label-free approach to quantify the relative abundance of intact proteins ranging from 0 to 30 kDa in two different states. In this chapter, we describe the integrated methodology for the large-scale profiling and quantitation of the intact proteome by liquid chromatography-mass spectrometry (LC-MS) without the need for metabolic or chemical labeling. This recent advance for quantitative top-down proteomics is best implemented with a robust and highly controlled sample preparation workflow before data acquisition on a high-resolution mass spectrometer, and the application of a hierarchical linear statistical model to account for the multiple levels of variance contained in quantitative proteomic comparisons of samples for basic and clinical research.Entities:
Keywords: Differential expression; Label-free quantitation; Proteoform; Quantitative mass spectrometry; Top-down proteomics; Top-down quantitation
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Year: 2016 PMID： 26867742 PMCID： PMC4777307 DOI： 10.1007/978-1-4939-3524-6_8
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745