Principle of TMT-Based Quantitative Proteomics Analysis

TMT (Tandem Mass Tags) quantitative proteomics analysis is a highly sensitive, multiplexed quantification technique based on mass spectrometry, widely used in proteomics research. This technology enables the relative quantification of multiple samples by labeling them with distinct tags and analyzing their intensities through mass spectrometry.

 

Structure of TMT Tags

TMT tags consist of three components: a reporter group, a balance group, and a reactive group. The reporter group generates characteristic fragment ions during mass spectrometry analysis, which can be precisely measured. The balance group compensates for the mass differences introduced by different tags, ensuring the comparability of signal intensities across samples. The reactive group covalently binds to the N-terminus of peptides, completing the labeling process.

 

1. Sample Labeling and Mass Spectrometry Analysis

In TMT quantitative proteomics analysis, samples from different sources are first enzymatically digested into peptides. These peptides are then labeled by reacting with the reactive group of the TMT tags. Each TMT tag has a distinct reporter group, which generates characteristic ions during mass spectrometry analysis. The labeled samples are combined and separated by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for detection.

 

2. Data Analysis and Quantification

During mass spectrometry, the reporter group of the TMT tag produces characteristic ion signals in the second stage of mass spectrometry (MS/MS), which are used for quantification. Relative quantification is achieved by comparing the signal intensities of reporter ions across different samples. Data analysis software such as Proteome Discoverer can automatically identify and integrate these signals, generating the corresponding quantitative data.

 

Advantages of TMT Quantitative Proteomics Analysis

Compared to traditional proteomics analysis methods, TMT quantitative proteomics offers advantages such as strong multiplexing capability, high sensitivity, and high throughput. These features have made it widely applicable in biomedical research, disease biomarker discovery, and drug target screening.

 

TMT quantitative proteomics analysis, with its unique tag design and advanced mass spectrometry technology, provides an efficient method for multiplexed sample quantification.