Unraveling mass transfer phenomena of large molecules in liquid chromatography: focus on biopharmaceuticals

Biopharmaceuticals are a class of emerging drugs that are obtained from or through living organisms. Biopharmaceuticals are large molecules with a complex composition, containing sugars, proteins, and/or nucleic acids. To ensure the stability, efficacy and safety of biopharmaceuticals, their composition needs to be accurately known. For this purpose, high-performance liquid chromatography (HPLC) can be used. HPLC typically separates analytes on a particle packed column, by flushing this column with a liquid with a tunable composition. When the analytes in a sample have different affinities for the stationary phase on the column and the liquid, they will migrate through the column with different velocities and separate. Many parameters affect the quality of this separation, by influencing the way the analytes are transferred through the column. These include structural column parameters, such as particle size, pore size, pore shape, and overall porosity of the column. Many studies have been performed to better understand the effect of these parameters on the separation of small molecules. However, this work
is currently lagging behind when it comes to large molecules, such as biopharmaceuticals. The main aim of this project therefore is to perform a detailed investigation of the parameters that influence the mass transfer of large molecules in HPLC, to better understand how HPLC columns can be designed to improve the separation of biopharmaceuticals in the future.