Mass Spectrometry Analysis Expert provides the latest proteomics, metabolomics information based on years of experience in this field.
Protein Data-independent Acquisition Quantitative Technology
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In the field of proteomics research, liquid chromatography-mass spectrometry (LC-MS/MS)-based discovery proteomics can detect and relatively quantify thousands of proteins in biological samples over the past decade or two. It has been widely used in various research, and reseaechers have developed two mainstream methods, label quantification (iTRAQ/TMT, SILAC) and label free, which are based on data-dependent acquisition (DADA). The method of collecting protein spectral data is also referred to as "shotgun".
However, the DDA data acquisition mode has inherent shortcomings such as poor repeatability, inaccurate quantification, large data loss, and difficulty in detecting low abundance proteins. MRM/SRM/PRM-based target proteomics can provide accurate absolute quantitative results for tens to hundreds of target proteins with high repeatability and is the gold standard for absolute quantitation of mass spectrometry. However, the flux is relatively low, and accurate quantitation of high-throughput proteins cannot be performed.
Is there a technology, which is not only of high-throughput, quantitative,but with accurate repeatability as well? The DIA technology (data-independent acquisition) developed and rapidly ignited in recent years is such the technology.
This technology combines the high-throughput and mass spectrometry of the traditional proteome shotgun with the quantitative and accurate advantages of the absolute quantitative gold standard MRM/PRM, which is named the most noteworthy technology of 2015 by Nature Methods. The number of published articles related to DIA has grown rapidly in the past three years and is the current fragrance in the field of proteomics research.
Protein DIA Quantitative Publication Trend (data source: Pubmed) DIA: DIA/SWATH The entire scanning range of the mass spectrum is divided into several parts, and all the parent ions in each part are selected, fragmented, and detected at high speed and cyclically, and all pieces of information of all the parent ions in the sample are obtained without any deviation.
Protein DIA Quantitative Analysis Workflow Applications of Protein DIA Quantitative Analysis: Medicine, botany, pharmacy, animal husbandry, food and environment.
As a commonly used technology in biological research today, mass spectrometry is becoming increasingly important in biological research by quickly and efficiently measuring the advantages of complex biological macromolecules and small molecules. Mechanism of M ass S pectrometry Mass spectrometry analysis is a method of analyzing a sample by measuring and analyzing the mass-to-charge ratio of the sample ions. The mass spectrometer is roughly divided into three parts , ion source, mass analyzer, and detector. The mass analyzer is the core component of the mass spectrometer, which determines the sensitivity, resolution, ability to generate ion fragments, and accuracy of the mass spectrometer. Before the sample is analyzed, it is first ionized in order to bring a certain charge to the sample molecule. Next, the charged ions will first driven through an accelerated electric field and fly into an analytical electric or magnetic field. Due ...
As a commonly used technology in biological research today, mass spectrometry is known as one of the most popular techniques in biological research by quickly and efficiently measuring the advantages of complex biological macromolecules and small molecules. The History of M ass S pectrometry The first mass spectrometer , invented by J. J. Thomson , has been around for more than 90 years . It was mainly used for the determination of inorganic elements in chemical experiments. With the development of mass spectrometry technology and computer technology, mass spectrometer has gradually been applied in the analysis of more complex biomolecules, including amino acid , protein, lipids, sugar and so on. Although the history of mass spectrometry and the time it has been applied to biological research are much shorter than those of microscopes, the importance of mass spectrometry in biological research is irreplaceable as a n accurate m...
Types of Commonly U sed M ass S pectrometer s The subjects of biological research are generally complex macromolecules, including proteins, nucleic acids, sugars, lipids, and various small molecules. The nature of various types of biomolecules also varies widely. Therefore, various types of mass spectrometers are used in biological research. The c ommon used ones for biological research include quadrupole mass spectromete r, t ime of f light m ass s pectrometer , ion trap mass spectrometer an d i on cyclotron resonance mass spectrometer . Time of Flight Mass Spectrometer (TOF) The mass-to-charge ratio of ions determined by this type of mass spectrometer is derived by analyzing the time of flight of the ions in the vacuum flight tube. After continuous improvement, the TOF mass spectrometer has greatly improved in resolution and precision. The accuracy can reach PPM level after the post-correction procedure, and the detected ion mass range...
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