Optimizing Aggregate Analysis – Pore Size

Introduction

Size exclusion chromatography (SEC) is one of the chromatographic separation modes used for large molecules such as proteins and polymers by their sizes. SEC is one of the crucial technique for characterizing protein-based therapeutic products. It is used for qualitative and quantitative analysis of fragments, monomers, dimers, and higher aggregates of therapeutic proteins. The variations in aggregates impact the applicability, efficacy, and safety of these therapeutic proteins. In SEC, smaller molecules enter the pores of the particle and proceed slowly along the axis direction of the column while molecules larger than the pore size are excluded from pores and elute at first from the column. In this application, we demonstrate the effect of pore size in SEC analysis. The analysis was performed using Shim-packTM Bio Diol columns with different pore sizes on a NexeraTM Bio UHPLC system.

October 22, 2020 GMT