Biopharmaceutical Disulfide Bond Analysis Service
Disulfide bonds are covalent linkages formed by the oxidation of thiol groups (–SH) between two cysteine residues. In protein molecules, disulfide bonds not only stabilize the spatial conformation but also directly determine protein folding and biological activity. For biopharmaceuticals such as monoclonal antibodies, recombinant proteins, and vaccines, correct disulfide bond pairing is a key factor in maintaining drug stability and biological function. Studies have shown that mispaired or broken disulfide bonds may cause conformational changes, increased aggregation, or even immunogenic risks, which seriously affect the safety and efficacy of drugs.
In drug development and quality control, Biopharmaceutical Disulfide Bond Analysis has become an indispensable characterization process. Through systematic disulfide bond analysis, the connectivity pattern of disulfide bonds in proteins can be revealed, as well as potential mispairing, breakage, or heterogeneity, thereby providing strong support for drug optimization, manufacturing consistency, and regulatory submission.
Figure 1. Protein Disulfide Bond Analysis Workflow
As a professional proteomics CRO, MtoZ Biolabs relies on high-resolution mass spectrometry platforms and well-established sample preparation workflows to provide a comprehensive and reliable biopharmaceutical disulfide bond analysis service that enables clients to gain accurate insights into protein structural integrity and stability. We are dedicated to helping academic and industrial clients systematically analyze disulfide bond characteristics, ensuring the structural integrity and clinical safety of biopharmaceuticals.
Services at MtoZ Biolabs
MtoZ Biolabs’ biopharmaceutical disulfide bond analysis service is based on advanced separation and mass spectrometry platforms, combined with diverse experimental strategies, to comprehensively analyze the connectivity and status of disulfide bonds in proteins.
1. LC-MS/MS peptide analysis under non-reducing conditions
Proteins are digested under non-reducing conditions to generate peptides that retain disulfide bonds. Using liquid chromatography–tandem mass spectrometry (LC-MS/MS), the cysteine residues linked by disulfide bonds can be directly analyzed. This method achieves site-level identification of disulfide bonds at the molecular level.
2. Comparative analysis of reduced and non-reduced samples
By comparing peptide mass spectrometry profiles under reducing and non-reducing conditions, the presence and location of disulfide bonds can be confirmed. This helps identify potential mispairing, free thiols, or missing disulfide bonds.
3. High-resolution mass spectrometry platform
With high-resolution mass spectrometers such as Orbitrap, subtle mass differences can be accurately distinguished, improving the analysis of complex disulfide bond patterns. Combined with multiple fragmentation modes (such as HCD and ETD), the coverage and accuracy of disulfide bond information are further enhanced.
4. Analysis at intact protein and fragment levels
In addition to peptide-level detection, disulfide bond analysis can also be performed at intact protein or functional fragment levels, providing a direct overview of molecular stability and disulfide bond distribution.
Analysis Workflow
1. Sample preparation: Receive purified biopharmaceutical samples from clients and perform pretreatment to ensure detection stability.
2. Enzymatic digestion: Digest under reducing or non-reducing conditions to generate peptides suitable for detection.
3. Mass spectrometry detection: Acquire disulfide bond-related information using high-resolution LC-MS/MS platforms.
4. Data analysis: Confirm disulfide bond connectivity based on databases and fragment ion features.
5. Result output: Generate a complete report of disulfide bond distribution and pairing.
Service Advantages
With advanced mass spectrometry platforms and a professional analytical team, MtoZ Biolabs provides a reliable and systematic biopharmaceutical disulfide bond analysis service that supports structural characterization, quality assurance, and regulatory compliance.
Contact us to obtain customized analytical solutions and make your biopharmaceutical development more efficient, precise, and compliant with international standards.