PRM Absolute Quantification: A Revolutionary Approach in Proteomics
Proteomics is an important means to study the composition and function of proteins in organisms. Over the past few decades, scientists have made many important breakthroughs through the continuous development of mass spectrometry technology. Among them, quantitative mass spectrometry technology has played a crucial role in proteomic research. PRM (Parallel Reaction Monitoring) absolute protein quantitation technology, as a new type of quantitative mass spectrometry method, provides a more accurate, sensitive, and high-throughput quantification method for proteomic research.
The PRM protein absolute quantitation technology is based on the high resolution and high sensitivity characteristics of the mass spectrometer, combining the advantages of selective reaction monitoring and parallel reaction monitoring. Its principle can be simply summarized into the following steps.
1. Sample Preparation
Extract and digest the test samples to obtain peptide mixtures.
2. Mass Spectrometer Setting
According to the peptide sequence of the target protein, design an appropriate mass spectrometry method, including the ion transmission parameters and collision energy of the mass spectrometer.
3. Data Collection
Data is collected through the mass spectrometer, recording the mass spectra of each peptide.
4. Data Analysis
Use professional mass spectrometry data analysis software to process the collected data and calculate the relative abundance of each peptide.
5. Standard Curve Plotting
Draw a standard curve through a series of standard samples with known concentrations for subsequent quantitative calculations.
6. Absolute Quantitation Calculation
Calculate the absolute abundance of the target protein based on the standard curve and the mass peak area of the test sample.
Advantages
Compared with traditional quantitative mass spectrometry methods, PRM protein absolute quantitation technology has the following notable advantages:
1. High Sensitivity
PRM technology utilizes the high resolution and high sensitivity of the mass spectrometer, which can detect low-abundance proteins, improving the accuracy of quantitation.
2. High Selectivity
Through selective reaction monitoring, PRM technology can quantitate the target peptides, reducing the impact of other interfering substances.
3. High Throughput
PRM technology can quantitate multiple peptides simultaneously, improving the efficiency and throughput of quantitation.
4. Accuracy and Repeatability
PRM technology can provide accurate and repeatable quantitation results through the establishment of standard curves and precise calculation of mass spectrometry data.
Application
PRM protein absolute quantitation technology has a wide range of applications in biomedical research. Below are some typical application fields:
1. Quantitation of Tumor Markers
PRM technology can be used to quantitate tumor markers, assisting in early diagnosis and treatment monitoring.
2. Pharmacokinetic Studies of Drugs
PRM technology can be used to quantitate drugs and their metabolites to study the metabolic pathways and kinetic characteristics of drugs.
3. Protein Interaction Studies
PRM technology can be used to quantitate protein interactions, revealing protein function and regulation mechanisms.
4. Discovery of Biological Markers
PRM technology can be used to screen and quantitate biological markers, providing new targets and methods for disease diagnosis and treatment.
As a new type of quantitative mass spectrometry method, PRM protein absolute quantitation technology provides a more accurate, sensitive, and high-throughput quantification method for proteomic research. Its advantages lie in its high sensitivity, high selectivity, high throughput, accuracy, and repeatability. In biomedical research, PRM technology has a wide range of application prospects and can play an important role in tumor marker quantification, pharmacokinetic studies of drugs, protein interaction studies, and discovery of biological markers. With the continuous development of mass spectrometry technology, it is believed that PRM protein absolute quantitation technology will play an even more important role in future research.
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