Radial scale-down of packed bed chromatography in a thin cylindrical tube for preparative media

Abstract

Microscale packed-bed chromatography columns (5 to 80. μL) were designed with cylindrical polyetheretherketone (PEEK) tubes for preparative ion exchange media by reducing internal column diameters with retained heights. Q Sepharose Fast Flow, soft agarose-based media, and POROS HQ, rigid polystyrene-based media, were packed in a thin column under the pressure of 0.3. MPa equally for various sizes of internal diameter ranging from 0.25 to 1. mm. The minimum internal diameter of tube was limited by the mean diameter of gel, thereby media could be packed in a plastic tube column that was no smaller than 5 times gel diameter, while maintaining comparable resin density. The microcolumn quantitatively showed equivalent performance when measured by total bound protein in breakthrough curve for both gels. Qualitative adsorption and desorption profiles showed that rigid gel exhibited slight increase in adsorption and separation efficiency as the column diameter decreased from 1. mm to 0.5. mm. The microscale PEEK tube column showed successful comparable performance with laboratory-scale liquid chromatography behavior, enabling it to offer a novel scale-down approach for the development of biopharmaceutical production process.