Three Fundamental Technologies of Proteomics

Proteomics, as an important research field in the field of biological products, focuses on the entire set of all proteins in the organism, helping us to deeply understand the molecular mechanisms in cells and organisms. In the research of proteomics, mass spectrometry analysis is one of the three basic techniques, which efficiently and accurately identifies protein samples, detects protein modifications and interactions, etc., providing a powerful means for deciphering proteomics.

 

Two-Dimensional Gel Electrophoresis

Two-dimensional gel electrophoresis is one of the earliest techniques used in proteomics. It separates protein samples according to their isoelectric point and molecular weight to form a two-dimensional protein map. In the first dimension, the protein samples are separated according to the charge of the isoelectric point, while in the second dimension, they are separated according to the molecular weight. By analyzing the two-dimensional gel electrophoresis map, the existence of proteins can be initially determined, protein changes can be detected, and potential protein markers can be discovered.

 

Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS)

Mass spectrometry technology is one of the most important technologies in proteomics, among which MALDI-TOF MS is a commonly used mass spectrometry method. This technique mixes protein samples with a matrix, then irradiates them with a laser, causing the protein and matrix to form characteristic peaks of charge-mass ratio together. Subsequently, these charged protein ions will produce differences in flight time based on their charge-mass ratio. By measuring the flight time and charge-mass ratio of these ions, the molecular weight of the protein can be determined. MALDI-TOF MS can rapidly identify protein samples in high throughput and is an important tool in proteomics.

 

Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

Liquid chromatography-tandem mass spectrometry is another important method in mass spectrometry technology. Compared with MALDI-TOF MS, LC-MS/MS can more accurately identify proteins in complex samples. This technique first separates the protein samples by liquid chromatography, and then sends the separated proteins into the tandem mass spectrometry for fragmentation. Subsequently, by measuring the charge-mass ratio of these protein fragments, the amino acid sequence of the original protein can be inferred, thus determining the identity of the protein. The high sensitivity and high resolution of LC-MS/MS make it an ideal choice for analyzing proteins in complex samples.

 

Through the above three basic techniques, proteomics can deeply reveal important information about the expression, structure, function, and interaction of proteins in organisms, providing strong support for research and medical applications in the field of biological products.