mAb aggregate detection – Analytical HIC as an Orthogonal Chromatographic Approach

Abstract

During the last decades, mAbs have proofed to be a very valuable medication for severe illnesses like auto immune diseases and cancer. However, to ensure a successful therapy and the least possible side effects, a thoroughly investigation of potential aggregates is crucial. The quality of aggregates can be diverse in terms of physico-chemical and physiological properties. Besides a declined therapeutic effect, mAb aggregates may also be immunogenic.

A detailed characterization of the different aggregate species requires resolution of the different species by an online analytical method, as aggregation is a dynamic process. Due to the rather hydrophobic nature of mAb aggregates, analytical HIC using 2.5 μm particles offers outstanding resolution of the different aggregates. Therefore, a targeted analysis of every single contained species is possible. Fluorescence detection and an applied light scattering device ensure maximum analysis sensitivity. Furthermore, we could show that the highly efficient non-porous resin allows a quantitative analysis, providing an actual back-up method for the verification of SEC results.