Evaluation of HPLC Systems for Monoclonal Antibody Analysis

This technical report evaluates modern and legacy high-performance liquid chromatography (HPLC) systems for size-exclusion analysis of aggregation and fragmentation in monoclonal antibody drugs. The study specifically compares the Alliance iS Bio HPLC System, Alliance e2695, and a competitive bio-inert HPLC system. The report outlines the challenges posed by extra-column dispersion and non-specific adsorption in traditional HPLC systems, which complicate data analysis due to band broadening and peak tailing. The Alliance iS Bio HPLC System is highlighted for its lower system dispersion and bio-inert flow path, which enhance chromatographic quality. Experimental results demonstrate that the Alliance iS Bio System provides improved peak resolution and accuracy in measuring low-molecular weight species compared to the other systems. The findings indicate that the Alliance iS Bio HPLC System is well-suited for bioanalytical applications, particularly in monitoring critical quality attributes of monoclonal antibodies throughout their lifecycle.