- Selection of proteases (trypsin, chymotrypsin, Lys-C, Glu-C) tailored to antibody type.
- Generation of peptides for optimal coverage and sequence verification.
Peptide Mapping based Antibody Sequence & Structure Analysis Service
Introduction
Peptide mapping is a cornerstone technique for comprehensive antibody sequence verification and structural assessment. It involves enzymatic digestion of the antibody followed by separation and identification of resulting peptides using high-resolution mass spectrometry. This approach enables confirmation of primary sequence, detection of post-translational modifications, identification of cleavage sites, and verification of disulfide bond connectivity.
Our Peptide Mapping based Antibody Sequence & Structure Analysis Service delivers detailed structural insights with high sensitivity and reproducibility. It is essential for confirming batch-to-batch consistency, validating expression and purification processes, and supporting development of therapeutic or research-grade antibodies.
Strategies
Proteolytic Digestion
Liquid Chromatography-Mass Spectrometry (LC-MS/MS)
- High-resolution separation and detection of peptide fragments.
- Identification of amino acid sequence, post-translational modifications (glycosylation, oxidation, deamidation), and truncations.
Disulfide Mapping
- Analysis under non-reducing conditions to identify correct disulfide pairing.
- Ensures correct folding and chain assembly critical for functional activity.
Quantitative Peptide Coverage Assessment
- Comparison of detected peptides with theoretical sequence.
- Identification of missing, modified, or unexpected peptides.
Integration with Structural and Functional Data
- Correlation with intact mass, SDS-PAGE, and BLI data for holistic structural verification.
- Supports informed decisions for process optimization and functional integrity assessment.
Required Materials
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Antibody Samples
- Concentration: ≥1 mg/mL; volume: 200–500 µL per digestion.
- Preferably free from high-concentration excipients that interfere with enzymatic digestion.
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Sequence Information
- Annotated primary sequence (FASTA or similar).
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Buffer Details
- pH, salts, stabilizers, and formulation notes.
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Optional Reference Standards
- Well-characterized antibody batch for method validation.
Publications
Peptide mapping is widely adopted for structural verification:
FAQs
What is the sequence coverage achievable?
Typically >95% for full-length IgG; coverage depends on protease selection and digestion efficiency.
Can peptide mapping detect PTMs?
Yes, including glycosylation, oxidation, deamidation, and truncations.
How does this method support structural verification?
Confirms primary sequence, chain integrity, disulfide connectivity, and highlights modifications affecting function.
Turnaround time?
12–18 business days, depending on sample complexity.
Are functional studies included?
Functional correlation is optional; BLI or ELISA can be integrated with mapping results.
Contact Us
Ensure the integrity of your antibody sequence and structural fidelity with our Peptide Mapping based Antibody Sequence & Structure Analysis Service. Our experts leverage enzymatic digestion, high-resolution LC-MS/MS, and disulfide mapping to deliver comprehensive sequence verification, post-translational modification identification, and structural confirmation.
Getting Started is Simple:
01 Initial Consultation
Contact our technical team to discuss your antibody characteristics, analytical goals, and any known challenges. We will advise on protease selection, digestion strategy, and analytical platform configuration.
02 Sample Submission
Provide purified antibody samples (≥1 mg/mL, 200–500 µL recommended), annotated sequence information (FASTA format), formulation details, and any reference standards. Correct labeling and cold shipment are critical to preserve integrity.
03 Customized Workflow Design
Our specialists design a tailored peptide mapping strategy to achieve maximum sequence coverage, PTM detection, and disulfide bond verification. Optional integration with functional assays can be arranged to correlate structure with activity.
04 Comprehensive Reporting
You will receive a detailed report with peptide coverage maps, PTM identification, disulfide connectivity, and quantitative analysis. Deviations from expected sequence or structural anomalies will be highlighted with actionable insights.
05 Ongoing Support & Follow-up
Our team can support batch-to-batch consistency studies, sequence confirmation of new variants, or further in-depth structural analyses as needed.
For project inquiries, sample submission, or tailored analytical planning, please reach out to our team. Provide your antibody sequence, formulation details, and any prior analytical data to ensure precise and high-confidence peptide mapping results.
For research use only. Not intended for any clinical use.
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