Optimizing Aggregates Analysis – Mobile Phase

Introduction

Protein-based therapeutic products such as monoclonal antibodies (mAbs) have shown to be effective against a variety of diseases. However, a slight difference or modification in the protein can alter its activity. Protein aggregation is one of the factors that causes the product unfit for use. Thus, accurate analysis of protein aggregates is required to ensure the applicability of therapeutic proteins. Size exclusion chromatography (SEC) is one of the chromatographic separation modes used for large molecules such as proteins and polymers by their sizes. It has been the main technique for analysis of fragments, monomers, dimers, and higher aggregates of therapeutic proteins. Despite being a simple assay, mobile phase condition is often needed to be optimized to improve peak shape and resolution of proteins. In this study, we describe analysis of trastuzumab, an anti-HER2 mAb using Shim-packTM Bio Diol column on Nexera TM Bio UHPLC. The effect of mobile phase composition (ionic strength and pH) on chromatographic separation of trastuzumab is presented in this application news.

June 9, 2020 GMT