Size exclusion chromatography (SEC) is an important tool for monitoring monomers, dimers, aggregates, and potential degradants in biotherapeutic protein samples, including monoclonal antibodies and their derivatives. As aggregation is considered a critical quality attribute, quantitation is required. This application note describes a simple and sensitive method for the quantitation of aggregates in a biotherapeutic mAb and antibody drug conjugate (ADC) using an Agilent AdvanceBio SEC 300Å, 7.8 × 300 mm, 2.7 µm column and an Agilent 1260 Infinity Bio-inert Quaternary LC. The method uses the same aqueous mobile phase, without the addition of organic modifier, for the analysis of the mAb and the more hydrophobic ADC. The optimized method was also able to monitor aggregate and degradants created by pH/temperature stress. This simple and reproducible method, coupled with the corrosion resistance of the instrument, is suitable for QA/QC analysis of mAbs and ADC in biopharma research.
Publication number: 5991-6303EN
Publication Date: March 23, 2016