Analyses of large proteins, antibodies, and modified proteins using a new series of advanced large pore HPLC materials

In this white paper, Advanced Materials Technology presents an overview with protein analysis data of its new HALO^® 1000 Å protein column series. The list of FDA-registered and commercialized therapeutic proteins continues to expand rapidly, including more than 400 biotherapeutics, such as enzymes, monoclonal antibodies (mAbs), antibody-drug conjugates (ADCs), and hybrid antigen binding structures. A key analytical approach for protein characterization remains reversed-phase HPLC (RPLC), due to the unique and informative capabilities of this method. Until very recently, researchers have used column technology for RPLC of intact mAbs, ADCs and their resulting fragments which has been fundamentally unchanged since the mid-1990s. Typically, such packing materials have consisted of 200 – 300 Å pore size, silica-based, fully porous particle (FPP) bonded phases. However, new, very wide, 1000 Å pore size superficially porous packings for RPLC of very large molecules have been developed to improve the resolution for, and the information gained from, higher resolution chromatographic separations of protein mixtures.