Comparison of Mass Spectrometry MRM With iTRAQ/TMT Quantification

In the field of biopharmaceutical research, protein quantification is an important task. Accurately measuring the expression levels of proteins is key to understanding disease mechanisms, drug development, and the discovery of biomarkers. In proteomics research, mass spectrometry is a common method. This article will focus on the two common proteomics quantification methods of mass spectrometry MRM (Multiple Reaction Monitoring) and iTRAQ/TMT quantification, and carry out comparative analysis.

 

Mass Spectrometry MRM Quantification

Mass spectrometry MRM is a quantitative method based on mass spectrometry technology. It achieves the quantification of target proteins by selecting specific peptide segments and transition reaction ion pairs. Firstly, the sample is analyzed by mass spectrometry, and the mass spectrum of peptides is obtained. Then, specific peptide segments and their corresponding transition reaction ion pairs are selected for monitoring. By measuring the peak area of these ion pairs, the content of target proteins can be inferred.

 

Mass spectrometry MRM quantification method features high sensitivity, high selectivity, and high throughput. It can monitor multiple peptide segments simultaneously, thus achieving quantification of multiple proteins. Additionally, the mass spectrometry MRM quantification method can also achieve absolute quantification by using stable isotope labeling technology (such as 15N labeling), improving the accuracy of quantification results.

 

iTRAQ/TMT Quantification

iTRAQ (Isobaric Tags for Relative and Absolute Quantitation) and TMT (Tandem Mass Tags) are two commonly used proteomics quantification methods. They introduce specific isotope-labeled reagents into samples to label proteins in different samples, then the labeled samples are mixed together for mass spectrometry analysis.

 

iTRAQ and TMT quantification methods have high throughput, high reproducibility, and high sensitivity. They can quantify multiple samples at the same time, thereby achieving high-throughput proteomics research. Additionally, iTRAQ and TMT quantification methods can achieve quantification accuracy by selectively labeling specific amino acid residues (such as arginine and lysine).

 

Comparison Between Mass Spectrometry MRM and iTRAQ/TMT Quantification

Both mass spectrometry MRM and iTRAQ/TMT quantification methods have important application value in proteomics research, but they also have some differences.

 

Firstly, mass spectrometry MRM quantification method has higher sensitivity and selectivity. It can selectively monitor target peptide segments, thereby avoiding the influence of other interfering substances in complex samples. In contrast, iTRAQ/TMT quantification methods require mass spectrometric analysis of the entire sample and may be affected by the complex sample matrix.

 

Secondly, mass spectrometry MRM quantification method is suitable for the quantification of a small number of target proteins. It can selectively monitor target peptide segments, thus achieving the quantification of a small number of target proteins. In contrast, iTRAQ/TMT quantification method is suitable for simultaneously quantifying multiple samples, but it may not be accurate enough for the quantification of a small number of target proteins.

 

Finally, mass spectrometry MRM quantification method can achieve absolute quantification, improving the accuracy of quantification results. By using stable isotope labeling technology, mass spectrometry MRM quantification results can be compared with standard products of known concentration to achieve absolute quantification. In contrast, iTRAQ/TMT quantification methods can only achieve relative quantification and cannot obtain absolute quantification results.

 

Mass spectrometry MRM and iTRAQ/TMT quantification methods are commonly used protein quantification methods in the field of biopharmaceutical research. Mass spectrometry MRM quantification method has the characteristics of high sensitivity, high selectivity, and high throughput, and is suitable for quantifying a small number of target proteins. iTRAQ/TMT quantification methods have the characteristics of high throughput, high reproducibility, and high sensitivity, and are suitable for simultaneous quantification of multiple samples. When choosing a quantification method, it is necessary to select based on specific experimental needs and sample characteristics.