Abstract: Lactoferrin is a glycoprotein with multiple biological activities, including antibacterial effects, immunomodulation, and promotion of iron absorption. It is widely used in high-end nutraceuticals and biomedicine. However, lactoferrin exists at very low levels in animal milk and coexists with a complex protein matrix, making its efficient and low-cost large-scale extraction and purification a key bottleneck in industrial development. This article briefly introduces the biological functions of lactoferrin, with a focus on the latest research progress in extraction processes and purification techniques. It provides an in-depth comparison of the practical applications and limitations of mainstream technologies such as ion exchange chromatography, affinity chromatography, and membrane separation. Additionally, the working principles of emerging advanced technologies, including reverse micelle extraction, magnetic nanoparticle separation, expanded bed adsorption, and precision fermentation, are reviewed. On this basis, commonly used quantitative methods and purity detection techniques for lactoferrin are outlined, and it is foreseen that the development of the lactoferrin industry will trend toward intelligent and green production models integrating multiple technologies.