Lactoferrin is one of the important nutraceutical with several physiological and biological functions. Bovine colostrum contains higher concentrations of lactoferrin than in the milk. The present work describes the use of two mixed mode sorbents, mercapto ethyl pyridine (MEP HyperCel™) and phenyl propyl amine (PPA HyperCel™), for the recovery of lactoferrin from bovine colostrum whey. The dynamic binding capacity study showed that, in MEP the mode of interaction of lactoferrin is similar to IgG and had a more or less same binding capacity of ~20 mg/mL of sorbent. In PPA, the interaction of lactoferrin was hydrophobic and was influenced by the addition of salt. The binding capacity of lactoferrin with PPA in absence and presence of salt was found to be ~17 and ~18 mg/mL sorbent respectively. The binding buffers used for the chromatographic experiments were 0.05 M sodium phosphate buffer, pH 7.4, with or without sodium chloride (0.15 M). A decreasing step pH gradient of pH 6.0, 5.5 and 4.0 was performed for elution. Both lactoferrin and immunoglobulin were obtained with high homogeneity. The recovered lactoferrin was confirmed by immunoblot analysis. The chromatographic elution fractions obtained from MEP Hypercel did not exhibit lactoperoxidase activity. The highest recovery of lactoferrin (~91%) with 2.9 fold rise in purity was obtained when the MEP resin column was used with the binding buffer without sodium chloride. Thus, MEP HyperCel™ could be a potential alternative to existing systems for separation of lactoferrin from bovine colostrum whey. © 2015, The Korean Society for Biotechnology and Bioengineering and Springer-Verlag Berlin Heidelberg.