MRM-Based Peptide Quantification Service

MRM-based peptide quantification is a targeted mass spectrometry quantification technique focused on the highly sensitive and specific quantification of known peptides. This service is based on a triple quadrupole mass spectrometry platform and is suitable for the precise quantification of low-abundance proteins or specific functional peptides in complex samples. Its principle involves the pairwise monitoring of precursor ions and specific product ions of target peptides at specific mass-to-charge ratios, thereby enabling highly selective quantitative detection.

 

MRM-based peptide quantification service is widely applied in protein expression level validation, biomarker verification, pharmacodynamic studies, signaling pathway monitoring, and quantitative analysis of clinical samples. It is particularly suitable for the accurate detection of low-abundance proteins in complex background samples. Compared to traditional quantification methods, MRM offers excellent reproducibility and high-throughput capabilities, making it suitable for parallel quantitative analysis of large-scale samples.

 



Figure 1. Schematic Diagram of MRM Scanning with Triple Quadrupole Mass Spectrometer.

 

Services at MtoZ Biolabs

Based on a highly sensitive mass spectrometry platform, MtoZ Biolabs offers MRM-based peptide quantification service focusing on the highly specific quantification of targeted peptides. This service is suitable for the precise quantification of known proteins or functional peptides in complex biological samples, covering various sample types such as recombinant proteins, plasma, and tissue extracts. Clients can obtain concentration changes of target peptides along with relative or absolute quantification results. The data exhibit strong reproducibility, making the service ideal for validating protein expression, screening candidate biomarkers, and conducting pharmacodynamic studies.

 

Analysis Workflow

1. Sample Preparation

The peptide solution is subjected to dilution, purification, or concentration to remove salts and interfering substances, ensuring stable mass spectrometry performance and consistent results.

 

2. LC-MS/MS Detection

High-performance mass spectrometry is employed in combination with nano or conventional liquid chromatography systems to perform MRM scans and capture characteristic peaks of the target peptides.

 

3. Data Analysis and Quantification

Specialized software (such as Skyline) is used to extract peak areas and fit standard curves, providing absolute or relative quantification values of the target peptides, along with assessments of reproducibility and accuracy.

 

4. Report Delivery

A comprehensive technical report is provided, including raw spectra, quantification results, QC analysis, and method parameters, supporting downstream biological or quality assessment studies.

 

Sample Submission Suggestions

1. Sample Type

It is recommended to provide peptide standards with known sequences or pre-synthesized target peptide solutions (purity ≥95% is preferred). Acceptable sample forms include aqueous solutions, buffers, or lyophilized powders.

 

2. Buffer Requirements

Avoid buffer systems containing high concentrations of salts, SDS, glycerol, DTT, or similar components, as they may interfere with mass spectrometry detection.

 

3. Sample Transportation

Samples should be stored at −80 °C and shipped on dry ice to ensure stability and integrity. Detailed guidelines for sample preparation and shipping are available to support standardized handling.

 

Service Advantages

1. High Specificity and Sensitivity

Utilizing the multiple reaction monitoring (MRM) mode, the method selectively detects specific peptides, significantly reducing background noise and enabling sensitive quantification of low-abundance proteins.

 

2. Excellent Reproducibility

Standardized mass spectrometry platforms ensure consistent and reliable results, making the service suitable for high-throughput analysis of large sample cohorts.

 

3. Customized Method Development

MtoZ Biolabs offers tailored experimental designs based on the client’s target proteins and research objectives to achieve accurate and focused detection.

 

4. One-Stop Analysis Service

The comprehensive workflow includes sample pretreatment, method development, mass spectrometry detection, and data analysis, streamlining experimental procedures and improving both research efficiency and data delivery quality.

 

Applications

1. Signaling Pathway Monitoring

MRM-based peptide quantification service enables the quantitative detection of key signaling peptides, supporting the analysis of activation states and functional changes within signaling pathways.

 

2. Biomarker Validation

Facilitates high-throughput verification of candidate peptides identified in earlier screening, improving the reliability and applicability of biomarker research.

 

3. Functional Peptide Research

MRM-based peptide quantification service can be used to evaluate the expression levels of functional peptides, supporting the investigation of peptide pharmacological mechanisms or biological regulatory pathways.

 

4. Quality Control and Process Monitoring

During peptide production, purification, and quality control, MRM quantification enables monitoring of specific peptide expression levels, ensuring product consistency and stability.

 

FAQ

Q1: Is it Necessary to Provide Information about the Target Peptide?

A1: It is recommended that clients provide the amino acid sequence of the target protein or peptide. If validated peptide information is available (e.g., from literature or preliminary experiments), it can greatly improve method development efficiency and quantification accuracy.

 

Q2: Does the Service Support Simultaneous Quantification of Multiple Peptides?

A2: Yes. MRM can simultaneously monitor multiple target peptides, making it suitable for multi-protein quantification. However, the number of monitored transitions should be balanced with sample complexity.

 

Q3: What Are the Advantages of MRM Compared to DIA and PRM?

A3: MRM offers superior specificity and sensitivity for high-throughput, accurate quantification of known targets. Compared to DIA, MRM enables simpler data processing; compared to PRM, MRM is more suitable for large-scale studies requiring higher transition capacity.

 

Deliverables

1. Comprehensive Experimental Details

2. Materials, Instruments, and Methods

3. Data Analysis, Preprocessing, and Estimation

4. Bioinformatics Analysis

5. Raw Data Files