PRM-Based Peptide Quantification Service

PRM (parallel reaction monitoring) is a targeted quantification technique based on high-resolution mass spectrometry, designed for highly sensitive and highly selective quantitative analysis of specific peptides. Compared to traditional MRM, PRM does not require preselection of product ion channels; instead, it simultaneously detects all fragment ions in a single scan, thereby improving data completeness and post-acquisition analysis flexibility. PRM is typically performed using high-resolution, high-accuracy mass spectrometry platforms such as orbitrap, in combination with either nano or conventional liquid chromatography systems.

 

PRM-based peptide quantification service is widely applied in validation experiments within proteomics research, including candidate biomarker verification, expression monitoring of key proteins in signaling pathways, drug target confirmation, functional peptide quantification, and post-translational modification analysis. It is particularly suitable for research scenarios that require highly accurate quantification of known targets.

 



Reyes, A V. et al. Frontiers in Plant Science, 2022.

Figure 1. Workflow of Parallel Reaction Monitoring.

 

Services at MtoZ Biolabs

Leveraging a high-resolution Orbitrap mass spectrometry platform, MtoZ Biolabs offers the PRM-based peptide quantification service focusing on precise quantification of predefined target peptides. This service integrates nano-liquid chromatography with high-resolution tandem mass spectrometry to comprehensively monitor all fragment ions of candidate peptides, enabling highly selective and sensitive quantification. It is suitable for targeted analysis of functional peptides, biomarker peptides, or modified peptides in complex samples such as plasma, tissues, and cells, providing clients with accurate and reliable expression profile data.

 

Analysis Workflow

1. Sample Preparation

After protein extraction, enzymatic digestion is performed to obtain target peptides. Purification or enrichment may be applied if necessary to enhance detection efficiency.

 

2. Target Peptide Selection and Method Development

Based on the target protein sequence, specific peptides are designed and high-response fragment ions are selected for subsequent PRM method setup.

 

3. High-Resolution Mass Spectrometry Detection

High-resolution mass spectrometry platforms such as Orbitrap are used to perform highly sensitive MS/MS scans on specific peptides, generating quantitative data.

 

4. Data Processing and Quantitative Analysis

Professional software is used for peak recognition, quantification ratio calculation, and statistical analysis, providing expression levels and variation trends of the target peptides.

 

Sample Submission Suggestions

1. Sample Types

A wide range of biological samples are supported, including serum, plasma, tissue homogenates, and pre-purified peptide solutions. Providing known sequences of target proteins or peptides is recommended to improve detection efficiency.

 

2. Sample Quality Requirements

Samples should be free from protein degradation and contamination by salts, lipids, or other impurities to ensure sensitivity and accuracy in mass spectrometry analysis.

 

3. Sample Storage and Transportation

Samples should be stored at −80 °C and transported on dry ice to avoid repeated freeze–thaw cycles. MtoZ Biolabs can assist clients with standardized procedures to ensure sample integrity.

 

Service Advantages

1. High Selectivity and Sensitivity

Based on high-resolution mass spectrometry platforms, PRM enables accurate identification and quantification of target peptides in complex backgrounds, especially suitable for low-abundance molecule analysis.

 

2. Excellent Quantitative Accuracy

Full-scan high-resolution MS/MS data acquisition allows precise and highly reproducible peptide quantification, making it suitable for rigorous scientific or validation studies.

 

3. Multiplexed Target Analysis

PRM supports simultaneous analysis of multiple target peptides, meeting the needs of multi-protein or multi-pathway research and improving experimental efficiency.

 

4. Customized Service

MtoZ Biolabs offers tailored experimental strategies based on client needs, supporting end-to-end customization from peptide selection to data analysis to facilitate precise research goals.

 

Applications

1. Signaling Pathway Research

PRM-based peptide quantification service can be used to quantitatively analyze the expression dynamics of key peptides in signaling pathways, elucidating regulatory mechanisms and activation states.

 

2. Biomarker Validation

Supports high-specificity validation of candidate peptides identified in preliminary screening, enhancing the reliability and applicability of potential biomarker research.

 

3. Exploration of Peptide Functional Mechanisms

PRM-based peptide quantification service enables expression quantification of functional peptides, revealing their roles in biological processes such as cellular regulation and metabolic modulation.

 

4. Quality Control and Sample Consistency Assessment

In biopharmaceutical development or process optimization, this service is used to monitor changes in specific peptide levels, ensuring sample quality and batch-to-batch stability.

 

FAQ

Q1: Can Multiple Peptides Be Detected Simultaneously?

A1: Yes. PRM supports the parallel detection of multiple peptides. However, it is recommended to keep the number of targets within a reasonable range to ensure sensitivity and data quality.

 

Q2: What Is the Difference between PRM and MRM?

A2: PRM offers higher selectivity and resolution, making it suitable for accurate detection in complex backgrounds. In contrast, MRM has advantages in throughput and speed, making it better suited for large-scale sample screening. PRM is ideal for projects requiring high-specificity validation.

 

Q3: Can Peptides with Post-Translational Modifications (PTMs) Be Analyzed?

A3: Yes. PRM technology has strong capabilities for identifying modification sites and is suitable for the quantitative analysis of PTM peptides such as phosphorylation, methylation, and acetylation.

 

Deliverables

1. Comprehensive Experimental Details

2. Materials, Instruments, and Methods

3. Data Analysis, Preprocessing, and Estimation

4. Bioinformatics Analysis

5. Raw Data Files