Antibody Characterization Tools
Analytical characterization of antibodies is critical during all stages of biologics drug development, from antibody purification to peptide mapping. Our solutions help you reduce hands-on time, scale easily, and generate reproducible data—backed by expert support.
Solutions for Antibody Characterization
Our reagents and ready-to-run assays can support you at every step of your antibody characterization workflow:
- Purify antibodies quickly from serum or cell culture using magnetic Protein A/G beads.
- Peptide mapping for sequence confirmation using mass-spec–grade trypsin, orthogonal proteases, or a low-pH digestion workflow.
- Post-translation modification (PTM) detection for glycosylation and artificial PTMs.
Peptide Mapping:
Tryptic Proteases
Peptide Mapping:
Orthogonal Proteases
Proteases for Peptide Mapping
Not sure which protease to use? Compare proteases in the table below to help you choose the right one for your purpose.
| Protease | Cleavage specificity | Primary purpose | Best when you need | Key advantages | Considerations |
|---|---|---|---|---|---|
| Trypsin Platinum | C-terminal of Lys (K) and Arg (R) | Gold-standard bottom-up peptide mapping | High specificity IDs; broad proteome/mAb coverage; compatibility with most LC-MS methods | Maximum digest specificity; high resistance to autoproteolysis | Can miss proline-adjacent sites; denaturant tolerance lower than Lys-C |
| Lys-C | C-terminal of Lys (K) | Pre-digest or standalone digest under harsh/denaturing conditions | Improved sequence coverage; tolerance to chaotropes; robust digestion of difficult proteins | Highly stable; great in challenging digestion conditions | Generates longer Lys-only peptides; follow with trypsin for classic peptides |
| Arg-C Ultra | C-terminal of Arg (R) | Minimize missed cleavages in trypsin workflows with Lys-C or Trypsin | Study PTMs on Lysine residues | Expands coverage with distinct cleavage chemistry | Arg-C Ultra is a cysteine protease and requires the presence of a reducing agent during digestion |
| ProAlanase | C-terminal of Pro (P) and Ala (A) | Access proline/alanine-rich regions; improve coverage of resistant sequences | Detecting sequence variants/PTMs in proline-rich segments | Complements trypsin/Lys-C; breaks tough regions | Requires acidic digestion conditions for optimal performance |
| IdeS | IgG hinge region (single site below hinge) | IgG subunit analysis and F(ab')2 prior to LC-MS | Middle-down or subunit mapping; glycoform/CQA assessment | Reproducible, antibody-specific cleavage for clean subunits | There is IgG subclass specificity |
Frequently-Asked Questions
What is peptide mapping and why is it important for monoclonal antibodies (mAbs)?
Peptide mapping digests a protein into peptides and identifies them by LC-MS to confirm sequence, detect variants, and localize PTMs. It’s a cornerstone of monoclonal antibody characterization and comparability studies.
What’s the difference between crude trypsin and mass spec grade trypsin?
Crude trypsin is low-purity and prone to autolysis, suitable for general protein digestion. MS-grade trypsin is highly purified and modified to reduce self-digestion, producing clean peptides for accurate mass spectrometry and proteomics analysis.
How do I choose Protein A vs. Protein G for purification?
Protein A binds human IgG1/IgG2/IgG4 strongly; Protein G performs better for some mouse and rat subclasses. Choose by species and subclass.
What is IdeS protease and how is it used?
IdeS is a highly specific protease that cleaves human IgG antibodies at the hinge region, generating F(ab’)₂ and Fc fragments. IdeS is favored for its rapid, precise, and complete IgG cleavage under mild conditions, making it a valuable tool in biotechnology.
Why are acidic protein digest conditions favored for antibodies?
Acidic protein digestion conditions are favored for antibodies because they preserve antibody structure, minimize unwanted cleavage or denaturation, and enhance specificity for enzymes like IdeS, ensuring efficient and controlled fragmentation for analysis.