Analysis of mAbs and Their Fragments by SEC and Orbitrap Mass Spectrometry
Overview
Size exclusion chromatography (SEC) columns are the ideal choice for monitoring aggregates of your monoclonal antibodies.
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.
Recommended SEC HPLC columns for aggregate analysis
HPLC column selection guide
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Resources
Importance of a Correct UHPLC Instrument Setup for Protein Aggregate Analysis by SEC
Lifetime Stability of SEC Columns for Protein Aggregate Analysis
A Universal Chromatography Method for Aggregate Analysis of mAbs
Specification Sheet: Acclaim SEC Columns
See Your Protein Therapeutics in High Resolution
Acclaim Column Selection Guide
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