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. Broader applications and value of PRM in protein quantification are summarized in Applications and Value of PRM Mass Spectrometry in Protein Quantification.

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.

Six-step PRM absolute quantification workflow from sample preparation and MS method setting through data analysis, standard curves, and absolute amount calculation

Figure 1. PRM absolute quantification proceeds from peptide preparation and selective acquisition through standard-curve calibration to absolute amount reporting.

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. How PRM supports low-abundance protein measurement is discussed further in How Does PRM Technology Facilitate the Analysis of Low-Abundance Proteins?.

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.

Four advantages of PRM absolute quantification including high sensitivity, selectivity, throughput, and standard-curve accuracy

Figure 2. PRM absolute quantification combines high sensitivity and selectivity with multiplex throughput and standard-curve–based repeatability.

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.

Four application areas of PRM absolute quantification including tumor markers, pharmacokinetics, protein interactions, and biomarker discovery

Figure 3. PRM absolute quantification is commonly applied to tumor marker measurement, pharmacokinetic studies, protein interaction assays, and biomarker discovery.

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. Researchers can also review the MtoZ Biolabs Parallel Reaction Monitoring (PRM) Service for project-specific absolute quantification panel design, calibration planning, and PRM analysis support.