SEC is a routine technique for the analysis of proteins, and SEC columns are often used to evaluate the aggregation profile of a protein. Protein aggregates are a product of degradation and must be closely monitored and kept within acceptable limits. This is a Critical Quality Attribute (QCA) for monoclonal antibodies (mAbs) and other antibodies because it can cause an immunogenic response in patients. Large antibody aggregates are sterically excluded from most pores and therefore flow quickly through an SEC column. Smaller proteins spend more time diffusing inside of the particle pores within the column’s stationary phase and take longer to elute. The smallest proteins, such as antibody fragments, can enter all pores and elute last. This ability to separate protein samples by size makes SEC.
✓ For high-resolution separation of mAbs including monomers, aggregates, and Fab & Fc fragments resulting from proteolysis
✓ For the separation of mAb conjugates from other compounds such as PEGylated analytes with a mass of 100 to 50,000 Daltons
✓ For the separation of mAb conjugates from other compounds such as PEGylated analytes with a mass of 1,000 to 1,000,000 Daltons
Analysis of mAbs and Their Fragments by SEC and Orbitrap Mass Spectrometry
Importance of a Correct UHPLC Instrument Setup for Protein Aggregate Analysis by SEC
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