HIC Analysis of Different Monoclonal Antibodies using Isopropanol as Modifier

Antibodies are often analysed in HIC mode because of the non-denaturating conditions used. Organic solvents in the mobile phase can cause the denaturation of antibodies. By adding of 15 % isopropanol the denaturation can be reduced and higher recoveries are achieved. The analysis of different MAbs using YMC’s BioPro HIC BF achieved high resolution with narrow peak widths.

This application note developed by the University of Geneva (Department of Analytical Pharmaceutical Chemistry) shows how to realise a robust chromatographic method using four commercially available MAbs: Adalimumab (Humira®), Brentuximab (commercially available as antibody drug conjugate “Brentuximab-vedotin” / Adcetris®), Pembrolizumab (Keytruda®) and Reslizumab (Cinqaero®) with 15 % isopropanol as modifier can be achieved with higher recoveries.