Method Development Strategies for Purity Analysis of Proteins by Intact and Subunit Analysis

A primary analytical technique for purity of recombinant proteins is reversed phase LC (RPLC). This simple, robust methodology has a relatively short analysis time, as well being high resolution, capable of separation of hydrophobic variants such as oxidation, glycoforms, and clipping. As such, it is common for monoclonal antibodies (mAbs), especially since it can be implemented for both intact and subunit levels without extensive method optimization.

This webinar will highlight method development for intact and subunit purity analysis for mAbs by RPLC. This will include best practices in method optimization, including how to best adjust method parameters such as temperature, gradient slope, and flow rate to obtain high-quality impurity profiling and ensure method robustness and reproducibility. Finally, we will also highlight how subunit analysis might apply to other mAb formats, including bispecific antibodies, IgG2/IgG4 Isotypes, and Fc-fusion proteins.

• Explore method development on a new wide pore C4 column chemistry to achieve faster run times and optimal separation

• Learn how to optimize temperature, flow rate, and gradients for mAbs and other mAb formats

• Understand how to achieve better reproducibility and resolution for intact RPLC impurity methods.