Improving Intact Biogeneric Protein Separations

Introduction

Aeris WIDEPORE is a recently introduced core-shell HPLC | UHPLC column specifi cally designed to provide improved resolution of intact proteins larger than 10 kilodaltons (KDa) in molecular weight. The improved resolution of proteins is accomplished by the use of a new core-shell particle morphology which minimizes Protein band-spreading that occurs during diffusion in and out of the coreshell particle. The result is narrower peaks and better resolution of closely eluting proteins. This improved resolution is especially useful for refolding, impurity, and post-translational modifi cation assays on intact biogeneric proteins where very slight differences between intact and modifi ed proteins elute closely on reversed Phase columns. Several examples are shown demonstrating the utility of Aeris WIDEPORE core-shell columns for such applications.

Since their debut over two years ago, Kinetex® core-shell columns have introduced a new paradigm in ultra-high performance by decoupling column effi ciency from high backpressures. This has resulted in small molecule applications with reduced run times and increased throughput without the need for expensive new UHPLC instrumentation. While analysis speed has some value for protein separations, the principal focus is more on improving resolution of proteins from near-identical post-translationally modifi ed impurities rather than reducing run times. A new wide pore core-shell column specifi cally designed to improve protein separations (Aeris WIDEPORE) has been introduced. Rather than utilize a similar morphology of small molecule core-shell columns with larger pores, a completely different particle that takes into account the slower diffusion of proteins in and out of porous particles was developed. A graphic representation of the Aeris particle is shown in Figure 1.