Comparison of iTRAQ/TMT Label-Based Quantitative Proteomics Techniques

Proteomics is a scientific field that studies the entire set of proteins and their functions within an organism. Understanding the changes in the proteome is crucial for understanding disease mechanisms, drug effects, and finding new therapeutic targets in the development of biological drugs. Among them, quantitative proteomics technology is one of the important means to study the changes in protein expression levels in the proteome. This article focuses on itraq and tmt labeling quantitative proteomics technology, and compares the two.

 

Itraq labeling Quantitative Proteomics Technology

itraq (isobaric tags for relative and absolute quantitation) labeling quantitative proteomics technology is a mass spectrometry-based quantitative method. It labels the peptides produced by protein digestion in different samples, and then quantifies them in mass spectrometry analysis. The itraq-labeled peptides produce specific ion peaks in mass spectrometry. By measuring the intensity of these ion peaks, the relative abundance of proteins in different samples can be inferred.

 

1. The Advantages of Itraq Labeling Quantitative Proteomics Technology

(1) High throughput

Itraq labeling can analyze multiple samples at the same time, improving experimental efficiency.

 

(2) High sensitivity

The ion peaks produced by itraq-labeled peptides in mass spectrometry are strong, which can detect low-abundance proteins.

 

(3) High accuracy

The accuracy of itraq labeling quantitative results is high, providing reliable data support.

 

2. Limitations of Itraq Labeling Quantitative Proteomics Technology

(1) Quantitative Error

The intensity of the ion peaks produced by itraq-labeled peptides in mass spectrometry is affected by various factors, such as ionization efficiency, mass spectrometer performance, etc., which may lead to quantitative errors.

 

(2) Inconsistent Labeling Efficiency

Peptides in different samples may have different labeling efficiencies, which can introduce quantitative bias.

 

(3) High Labeling Cost

The price of itraq labeling reagents is high, which is costly for large-scale experiments.

 

TMT Labeling Quantitative Proteomics Technology

tmt (tandem mass tag) labeling quantitative proteomics technology is also a mass spectrometry-based quantitative method. Similar to itraq labeling, tmt labeling also labels peptides in different samples and then quantifies them in mass spectrometry analysis. Tmt-labeled peptides produce different mass-to-charge ratios in mass spectrometry. By measuring the intensity of these mass-to-charge ratios, the relative abundance of proteins in different samples can be inferred.

 

1. The Advantages of TMT Labeling Quantitative Proteomics Technology

(1) Multiplex Labeling

TMT labeling can analyze multiple samples at the same time, improving experimental efficiency.

 

(2) High Accuracy

The accuracy of tmt labeling quantitative results is high, providing reliable data support.

 

(3) Consistent Labeling Efficiency

The labeling efficiency of tmt labeling reagents is relatively stable, reducing quantitative bias.

 

2. Limitations of TMT Labeling Quantitative Proteomics Technology

(1) Large Labeling Substance

The molecular weight of tmt labeling reagents is large, which may affect the sensitivity of mass spectrometry analysis.

 

(2) Expensive Labeling Substance

The price of tmt labeling reagents is high, which is costly for large-scale experiments.

 

Comparison of Itraq and TMT Labeling Quantitative Proteomics Technology

itraq and tmt labeling quantitative proteomics technologies are similar in principle, both labeling peptides for quantitative analysis. Their differences are mainly reflected in the following aspects:

 

1. Labeling Efficiency

the intensity of the ion peaks produced by itraq-labeled peptides in mass spectrometry may be affected by labeling efficiency, while the labeling efficiency of tmt-labeled peptides is relatively stable, reducing quantitative bias.

 

2. Price

the price of itraq labeling reagents is relatively lower, while the price of tmt labeling reagents is higher. Therefore, itraq labeling technology is more economical in large-scale experiments.

 

3. Analysis Throughput

itraq labeling can analyze multiple samples at the same time, improving experimental efficiency, while tmt labeling can analyze more samples at the same time, further improving experimental efficiency.

 

Both itraq and tmt labeling quantitative proteomics technologies are important quantitative proteomics methods. The selection of which technology to use should be based on experimental requirements, budget, and sample quantity. Regardless of which technology is chosen, the development of quantitative proteomics technology will provide more information and support for the development of biological drugs, promoting the progress of drug research.