Analysis of protein drugs aggregation Using Size Exclusion Chromatography

User Benefits

- Improving the separation of protein drugs and those aggregates and/or decompositions by suppressing secondary interactions such as electrostatic or hydrophobic interaction. - Highly reproducible data can be acquired even when using a mobile phase containing high concentrations of corrosive salts.

Introduction

Therapeutic proteins such as monoclonal antibodies (mAbs) and antibody drug conjugates (ADCs) easily aggregate due to changes in temperature, pH, concentration and so on. Aggregation in the production of mAb and ADC negatively affects their efficacy and safety. Therefore, the amount of aggregation should be monitored. Size exclusion chromatography (SEC), which separates molecules by differences in size, is one of the most utilized analytical methods for the detection of aggregation in therapeutic proteins,. It is known that adsorption occurs due to electrostatic interactions between mAb and stationary phases, and adsorption occurs due to hydrophobic interactions between ADC and stationary phases. These secondary interactions cause poor separation of aggregates, monomers, and fragments. Hence, it is necessary to consider these secondary interactions to ensure the reliability of data acquisition. This article describes an aggregate analysis using a Shimadzu “Shim-pack^TM Bio Diol” , size exclusion chromatography column with “Nexera XS inert”, an ultra high performance liquid chromatograph. This chromatograph has high salt tolerance and metal-free flow path and allows to use highly salted mobile phases and prevents sample adsorption.

February 2, 2022 GMT