- Complete acid hydrolysis of antibody samples into free amino acids.
- Quantitative separation using high-performance liquid chromatography (HPLC) with pre- or post-column derivatization for detection.
- Determination of individual amino acid mole ratios relative to expected sequence.
- Detection of amino acid anomalies indicative of misincorporation, truncation, or degradation.
Amino Acid Composition Analysis based Antibody Structure Verification Service
Introduction
Accurate knowledge of antibody primary structure is foundational for understanding its functionality, stability, and manufacturing consistency. Even minor deviations in amino acid composition can affect antigen binding, immunogenicity, and shelf-life. Amino acid composition analysis provides an essential verification of the expected sequence, complementing peptide mapping, intact mass analysis, and functional assays. This method detects unexpected substitutions, truncations, or post-translational modifications that may not be immediately apparent from SDS-PAGE, Western blot, or routine chromatographic assays.
Our Amino Acid Composition Analysis based Antibody Structure Verification Service delivers a quantitative and comprehensive assessment of the amino acid content of antibodies. We provide precise mole-per-mole analysis to confirm that the antibody conforms to its theoretical primary structure, ensuring consistency with design specifications and supporting quality control, process development, and research applications.
Strategies
Hydrolysis-Based Amino Acid Analysis
Mass Spectrometry-Based Composition Verification
- High-resolution LC-MS/MS for peptide-level confirmation of amino acid sequences.
- Quantification of residues, glycosylation sites, and oxidation-prone residues.
- Comparison against theoretical amino acid counts derived from sequence data.
Orthogonal Verification
- Comparison of hydrolysis-based results and MS-derived data to ensure consistency.
- Integration with intact mass analysis to confirm molecular weight alignment with verified amino acid composition.
Interpretation and Quality Insights
- Deviations from theoretical composition are highlighted and interpreted in the context of production process, expression system, or storage conditions.
- Reports include recommendations for purification optimization or additional verification if discrepancies are observed.
This multi-platform strategy ensures high confidence in amino acid-level integrity, supporting downstream functional and structural studies.
Required Materials
To conduct a robust amino acid composition analysis, clients should provide:
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Antibody Material
- Purified antibody at ≥ 1 mg/mL.
- Minimum volume: 200 µL for standard hydrolysis and MS analyses.
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Sequence Data
- FASTA or annotated sequence for comparison with measured composition.
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Buffer Information
- Details on formulation buffer, salts, stabilizers, and preservatives.
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Optional Controls
- Reference antibodies or peptides for method validation.
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Project Notes
- Specific areas of concern, such as suspected truncation or modification, to guide focused analysis.
Proper labeling and cold shipment are recommended to maintain sample integrity.
Publications
Amino acid composition analysis has long-standing use in protein characterization and is widely referenced in the literature:
These publications support the use of amino acid analysis as a critical verification step for antibody structural fidelity.
FAQs
How does amino acid composition analysis complement peptide mapping?
Peptide mapping provides sequence-specific identification, while composition analysis confirms overall residue counts, detecting anomalies that may be missed at the peptide level.
Can modified residues be detected?
Yes. Hydrolysis-based analysis can detect residues like methionine sulfoxide indirectly, while MS-based approaches can directly identify PTMs such as oxidation, deamidation, or glycosylation.
How long does the analysis take?
Standard turnaround is 10–14 business days, depending on sample number and complexity.
What sample quality is required?
High-purity antibodies reduce interference and improve accuracy; formulations with high excipient content may require buffer exchange.
Are results quantitative?
Yes. Both hydrolysis and MS methods provide quantitative mole ratios of amino acids compared to theoretical sequence composition.
Contact Us
Ensure the fidelity of your antibody sequence with our Amino Acid Composition Analysis based Antibody Structure Verification Service. Our experts integrate hydrolysis-based HPLC, LC-MS/MS, and orthogonal verification to deliver accurate and actionable insights into antibody structure.
For sample submission, project consultation, or customized service planning, please contact our team. Provide antibody sequence data, concentration, and formulation information for optimized analysis planning.
For research use only. Not intended for any clinical use.
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