Strategies for the Chromatographic Purification of Lactoferrin from Dairy Sources and Fermentation Broth

Abstract

In this study, we compare the effectiveness of two distinct resins for the purification of lactoferrin (LF) from skimmed milk and fermentation broth. The first resin, a highly porous and soft agarose-based material, exhibits strong hydrophilic properties, while the second resin, composed of rigid polymethacrylate, is more hydrophobic. The agarose-based resin, SP Seplife 6AG XL 200, demonstrates superior performance in purifying bovine lactoferrin (bLF) from milk and whey. Its hydrophilic structure minimizes undesired hydrophobic interactions, achieving a purity of 97% and a yield of 90%. Notably, the agarose resin effectively isolates lactoferrin despite its low concentration (<0.3%) and the presence of a similar protein, lactoperoxidase, which tends to coelute. Conversely, for the purification of recombinant bLF (rbLF) from the fermentation broth, the Seplife LXPM SP 5504C resin outperforms in both yield and purity, reaching 99% and 98%, respectively. This resin facilitates elution using only 0.5 M NaCl—an improvement over the 0.7 M NaCl required for the agarose resin. Both agarose and methacrylic resins excel at purifying feeds with high concentrations of rbLF (>10 mg/mL), while consistently maintaining excellent purity levels. Both resins are designed to be cost-effective and scalable and can be used in different chromatography systems employing axial or radial flow columns. Overall, the choice of resin significantly influences the purity and yield of lactoferrin, highlighting the importance of resin selection based on the source of protein.