Is Labeling Required for 4D-DIA Quantitative Proteomics
4D-DIA (Four-Dimensional Data-Independent Acquisition) is an innovative proteomics technique designed for comprehensive quantitative protein analysis. This method synergizes four dimensions of information: time, ion mobility, mass, and intensity, thereby enhancing the accuracy and efficiency of protein identification and quantification.
In the context of 4D-DIA proteomics, conventional chemical labeling methods such as iTRAQ or TMT are generally not necessary for quantification. The critical steps and characteristics of 4D-DIA quantitative proteomics include:
Sample Preparation
1. Protein Extraction
Proteins are isolated from biological samples.
2. Protein Digestion
Proteins are enzymatically digested into peptides using agents like trypsin.
Liquid Chromatography-Mass Spectrometry (LC-MS/MS)
1. Chromatographic Separation
Peptides are separated using liquid chromatography (LC).
2. Ionization
The separated peptides undergo electrospray ionization (ESI) to become charged ions.
3. Mass Spectrometry Detection
A mass spectrometer (MS) conducts a mass analysis on the peptides.
Ion Mobility Separation (IMS)
Prior to mass spectrometry, ion mobility separation (IMS) categorizes peptides based on their shape and charge.
Data Acquisition and Analysis
1. Data-Independent Acquisition (DIA)
DIA captures comprehensive fragment ion data for all peptides simultaneously, unlike methods targeting specific ions.
2. Quantitative Analysis
Bioinformatics processes mass spectrometry data to identify and quantify proteins. The holistic ion data provided by DIA facilitates relative quantification across multiple samples without the need for chemical labeling.
Advantages of 4D-DIA Technology
1. High Throughput and Sensitivity
Capable of simultaneous analysis of numerous peptides, it is ideal for expansive proteomic research.
2. Label-Free Quantification
Relies on direct comparison of mass spectrometry signal intensities to achieve relative quantification, eliminating the need for chemical labeling.
3. Enhanced Reproducibility and Coverage
The data-independent approach ensures high reproducibility and extensive proteome coverage.
4D-DIA quantitative proteomics negates the necessity for traditional chemical labeling, achieving precise protein quantification and identification through advanced mass spectrometry data collection and bioinformatics analysis.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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