Antibody Characterization Guide: Sample Preparation, Analytical Methods, and Data Interpretation
- Confirm sequences by MS or cDNA when needed.
- SEC-HPLC and CE-SDS quantify aggregates and fragments.
- Glycan and PTM mapping affect Fc function.
- SPR/BLI measures binding kinetics.
- DSC and DLS assess stability.
Systematic antibody characterization documents sequence, purity, modifications, binding, and stability before research or therapeutic use.
Key Takeaways
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Core Workflow
Sequence confirmation, purity analysis, glycosylation/PTM mapping, binding assays, and stability testing form the standard path.
Challenges
Combine SEC-HPLC with DLS for aggregation; use LC-MS/MS for PTMs; de novo sequencing for unknown commercial antibodies.
Conclusion
Structured characterization reduces failed experiments and supports confident antibody programs.
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