Differential Analysis of Proteome

Proteomics is a field of study that investigates all proteins and their interactions within a biological organism. Among this, differential analysis of the proteome is a significant branch of proteomics, aiming to compare the differences in protein expression between different biological samples, thereby revealing biological processes and disease mechanisms. There are mainly two methods for the differential analysis of the proteome: One is to screen differential proteins through two-dimensional electrophoresis technology, and then perform mass spectrometry analysis to identify information related to differential proteins; the other is a high-throughput sequencing technology based entirely on mass spectrometers, which finds differential proteins by quantitatively analyzing protein peptides after labeling.

 

Two-Dimensional Gel Electrophoresis Technology (2-DE)

2-DE is the earliest technology applied in proteomics research. Up to now, it is still a common separation method in proteomics research. The core of proteomics research is to ensure that the separated proteins have high resolution and reproducibility. 2-DE, with its capability of separating thousands of proteins while quantitatively analyzing differentially expressed proteins through staining technology, is widely used in proteomics research. The main steps include: protein sample preparation, isoelectric focusing (IEF), SDS-PAGE, staining, etc.

 

Mass Spectrometry (MS)

In the entire proteomics research, accurate identification of separated proteins is the most critical step. Mass spectrometry identification technology has high sensitivity, high throughput, and high timeliness, playing an important role in proteomics research. Traditional mass spectrometry technology can only be used for the analysis of small molecular weight substances, but with the advancement of technology, mass spectrometry technology has also made considerable progress, being able to detect more complex substances with higher automation and accuracy. Common protein mass spectrometry identification technologies include: matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), electrospray mass spectrometry (ESI-MS), protein fingerprinting identification (PMF), tandem mass spectrometry (MS/MS), liquid chromatography-mass spectrometry (LC-MS) and so on.