Multiple protein quantification method based on equiponderous dimethylation labeling

Abstract

The invention relates to a proteome multiple quantification method. With use of a property that a dimethylation reaction has different rates on a peptide fragment N tail end amino and a peptide fragment C tail end lysine side chain amino under acidic conditions, dimethylation labeling of an N tail end and a C tail end of a peptide fragment is realized successively under acidic and alkaline conditions. Various isotope forms of a dimethylation labeled reagent are organically combined to realize multiple labeling of the peptide fragment sample. The first-level spectrum mass-to-charge ratios of the peptide fragment after multiple labeling are exactly the same, the mass-to-charge ratios of second-level spectrum fragment ions have differences, and the corresponding second-level spectrum fragment ion intensity is extracted for multiple quantitative analysis. The method has the advantages of high quantitative accuracy, good precision and wide dynamic range, can perform simultaneous quantitative analysis of sixfold protein samples at the same time, greatly improves the flux of protein quantitative analysis, and saves the analysis time.

Description

technical field [0001] The invention relates to an equal-weight multiple-label quantification method, which can realize simultaneous large-scale quantification of up to six protein samples. The method has the advantages of high labeling efficiency, good labeling selectivity, high quantitative accuracy, precision, coverage, wide dynamic range, and high throughput of proteome quantitative analysis. It can be used for quantitative proteome analysis of large sample volume biological samples and dynamic processes of organisms. Background technique [0002] Changes in protein content can reflect life processes and reveal disease states, and exploring the changes can provide basic data support for finding drug targets and potential disease markers. In view of the large number of proteome samples and the content changes with time and space, it is urgent to develop quantitative methods that can simultaneously analyze multiple proteome samples on the premise of ensuring quantitative ...

AI Technical Summary

Problems solved by technology

Among the quantitative methods based on secondary spectrum, the quantitative method based on reporter ions can complete iTRAQ eight-fold labeling (Proteomics, 2007, 7, 3651.), TMT six-fold labeling (Anal Chem, 2008, 80, 2921.), but there is an underestimation effect, affecting quantitative accuracy

Recently, quantitative methods based on fragment ions have attracted more and more attention. At present, there are IPTL triple labeling (Anal.Chem.2013, 85, 2478.), mass loss quadruple labeling (Anal.Chem.2013, 85, 10658.), This method has high quantitative accuracy, good precision, and wider dynamic range, but the current throughput is small

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