Protein Disulfide Bond Analysis Service
Based on the high-resolution mass spectrometry (LC-MS/MS) platform and liquid chromatography (LC) system, MtoZ Biolabs has launched the protein disulfide bond analysis service which can perform systematic detection and quantitative analysis of disulfide bond connection patterns, positional distribution, and structural integrity in protein samples. This service combines control experiments under reducing/non-reducing conditions with optimized digestion workflows to reveal the connection modes of disulfide bonds and evaluate their impact on protein conformational stability and functional activity. The final output data include disulfide bond site identification results, modification distribution characteristics, and related functional annotations, providing reliable support for researchers in structural analysis, functional validation, and quality control. MtoZ Biolabs provides services including but not limited to the following:
1. Qualitative and Quantitative Analysis of Disulfide Bonds
Accurately identify and measure the presence and abundance changes of disulfide bonds through high-resolution mass spectrometry, providing comprehensive structural information.
2. Topological Analysis of Disulfide Bonds
Using non-reducing enzymatic digestion and mass spectrometry analysis, we comprehensively reveal the pairing patterns of disulfide bonds within proteins.
3. Dynamic Disulfide Bond Analysis
Monitor the formation and cleavage processes of disulfide bonds under different conditions or environmental stimuli, evaluating their dynamic changes in functional regulation.
Overview
Protein disulfide bond is a covalent linkage formed by the oxidation of sulfhydryl groups from two cysteine residues, and it is a key factor in maintaining protein conformational stability and functional activity. Disulfide bonds not only participate in protein folding but also play important roles in regulating molecular stability, signal transduction, and metabolic processes. Protein disulfide bond analysis has been widely applied in biopharmaceutical quality control, verification of structural integrity of protein drugs, complex proteome studies, and the investigation of protein functional regulation under environmental stress conditions, providing reliable structural and functional data support for both scientific research and industry.
Ren T, et al. Biotechnol Bioeng. 2021.
Figure 1. The Structure of Disulfide Bond.
Services at MtoZ Biolabs
1. Target Protein Disulfide Bond Analysis
MtoZ Biolabs can perform disulfide bond structural characterization for specific target proteins, identifying correct pairing patterns and any abnormal linkages. Leveraging high-resolution LC-MS/MS, we can reveal the impact of disulfide bonds on protein folding, stability, and function.
2. Disulfide Bond Proteomics Analysis
MtoZ Biolabs integrates targeted enrichment strategies with large-scale mass spectrometry to systematically identify disulfide bond features of multiple proteins in complex samples and evaluate structural changes under different treatment conditions. This analysis supports the elucidation of redox-regulated networks and disease-related protein structural remodeling trends.
Analysis Workflow
1. Sample Preparation
Extract sample proteins and process them under reducing/non-reducing conditions to ensure sample stability.
2. Protein Digestion
Perform enzymatic digestion under non-reducing conditions to retain disulfide bond information, with parallel reducing digestion as a control.
3. Peptide Enrichment
Enrich disulfide bond-containing peptides through chemical capture or selective separation to improve detection rate.
4. Mass Spectrometry Detection
Based on LC-MS/MS combined with liquid chromatography separation, precisely identify and quantify disulfide bond patterns and distribution.
5. Data Analysis
Database comparison outputs site information, pairing patterns, and functional annotation to generate a complete disulfide bond map.
Figure 2. The Workflow of Protein Disulfide Bond Analysis.
Sample Submission Suggestions
1. Sample Type and Quantity
2. Sample Storage
It is recommended to store samples at -80°C for long-term preservation and to avoid repeated freeze-thaw cycles in order to minimize protein degradation or disulfide bond breakage.
3. Sample Transportation
Samples should be transported on dry ice or under cold chain conditions, and sealed containers should be used to ensure sample integrity and the stability of disulfide bonds during transportation.
Service Advantages
1. High-Resolution Detection
Relying on the LC-MS/MS platform, disulfide bond information can be accurately captured in complex samples.
2. Combination of Multiple Methods
By combining reducing/non-reducing controls, enrichment, and multiple digestion strategies, connection patterns are validated from multiple perspectives.
3. One-Stop Service
From sample preparation to data analysis, complete workflow support is provided.
4. Customized Solutions
According to experimental objectives and sample characteristics, analysis strategies are flexibly adjusted to meet diverse needs.
Applications
1. Protein Folding Studies
Protein disulfide bond analysis service can be used to explore the role of disulfide bonds in protein folding and refolding processes.
2. Quality Control
In the detection of antibodies, recombinant proteins, and other bioproducts, the disulfide bond state is an important indicator of stability and consistency.
3. Cellular Function Studies
It can monitor changes in disulfide bonds during signaling and cellular responses to analyze regulatory mechanisms.
4. Environmental Stress Studies
Evaluate changes in disulfide bonds under oxidative stress or environmental variations to reveal cellular adaptation and maintenance of homeostasis.
Deliverables
1. Comprehensive Experimental Details
2. Materials, Instruments, and Methods
3. Identification and Quantification of Disulfide Bonds
4. Topological Analysis and Functional Annotation
5. Bioinformatics Analysis
6. Raw Data Files