Lactoferrin is a multifunctional 80kDa iron-binding, sialylated glycoprotein of the transferrin family, composed of two homologous lobes, each capable of binding one Fe³⁺ ion. Abundant in human milk, lactoferrin contributes to iron homeostasis, antimicrobial defence, immune modulation, and intestinal development [1, 2]. However, its isolation from bovine milk presents a significant challenge for the dairy industry. Increased global demand for lactoferrin drives the need for efficient ion-exchange processes to enhance yield without compromising purity. We investigated the impact of processing parameters and elution conditions on the yield and purity of lactoferrin using Sunresin SP Seplife®, an alternative commercial resin, at the laboratory scale. Two gradient elution methods and a step elution method were evaluated with pasteurised milk using an AKTA Pure 25M Fast Protein Liquid Chromatography system. We demonstrated that gradient methods 1 and 2 in pasteurised skim milk, achieved lactoferrin yields of 78.87mg/L and 79.74mg/L, respectively, representing 18-19% higher recovery than step elution (66.93mg/L). Overall, the gradient elution methods provided a feasible alternative to step elution, offering improved yield while maintaining lactoferrin purity above 90% across the treatments. These results provide an innovative and sustainable ion-exchange chromatography approach for enhanced recovery of high-value lactoferrin, thus improving resource utilisation and supporting the production of functional food ingredients.