Efficient Method Development of Monoclonal Antibody Size Variants by Size Exclusion Chromatography

User Benefits

- The separation between monoclonal antibody aggregates, monomers, and fragments can be visualized by design space to efficiently find the optimal condition that provides enough resolution as well as robustness. - Nexera™ XS inert (UHPLC system) offers high resistance to corrosion for stable data acquisition even when using mobile phases with high salt concentration. - pHM-40 not only provides real-time monitoring of the mobile phase pH but also visualizes the pH profile together with the chromatogram.

Introduction

Antibody drugs using monoclonal antibodies (mAbs) poseconcerns about forming aggregates during production andstorage in terms of their impact on safety and efficacy. ICH-Q6Brequires the separation of impurities such as aggregates andfragments in antibody drugs and to determine their content.Therefore, monitoring these impurities by size-exclusionchromatography (SEC) is one of the most important analysesduring the production of mAb. For SEC analysis, It is required to optimize the separationbetween aggregate, monomer, and fragments by changing saltconcentration and mobile phase pH while considering theinteraction between mAbs and column packing materials. Onthe other hand, using the mobile phases with high saltconcentration can cause damage on LC systems. This articledescribes an example of efficient method optimization forproviding enough separation and robustness for mAb sizevariants by using Nexera XS inert that is highly corrosion-resistant, pH monitor (pHM-40), and LabSolutions MD (adedicated software for supporting method development).

September 14, 2023 GMT