The biopharmaceutical industry continues to develop protein-based biotherapeutics in increasing numbers. Due to their complexity and biotechnological production, there are many attributes that need to be analyzed to guarantee their safety and efficacy. Peptide mapping is used to measure several critical quality attributes (CQA) required for the characterization of any biotherapeutic protein. The analysis is used to confirm that the correct sequence has been expressed for the protein and to check for post-translational and chemical modifications.
This study shows a fully automated UHPLC setup applying a simple and efficient online solid phase extraction method. Showcase assay performance by using a column cartridge for sample trapping and a Thermo Scientific™ MAbPac™ RP column for separation with water/acetonitrile-based gradients. Leverage the new technologies such as the Thermo Scientific™ Vanquish™ UHPLC platform in combination with the Thermo Scientific™ Q Exactive™ HF Hybrid Quadrupole Orbitrap mass spectrometer.
The Thermo Scientific Dionex UltiMate 3000 LC Bio-LC system is applied for the Hydrophobic Interaction (HIC) analysis of Rituximab. The separation was performed on a Thermo Scientific MabPac HIC-20 column at UV detection of 214 nm. With a total run time of 38 minutes the method allows separation of all oxidized and deamidated impurities of Rituximab.
The Thermo Scientific Dionex UltiMate 3000 Bio-LC system is applied for the reverse phase analysis of Rituximab (Drug Substance sample). The separation was performed on a Thermo Scientific MAbPac RP column. with UV detection at 214 nm. With a total run time of 70 minutes the method allows separation of all oxidized and deamidated impurities of Rituximab
The Thermo Scientific Dionex UltiMate 3000 LC Bio-LC system is applied for the Size exclusion analysis of Rituximab. The separation was performed on a Thermo Scientific MabPac SEC-1 column with a UV detection at 220 nm. With a run time of 30 minutes, the method allows easy separation of Aggregates and Dimer of Rituximab
