Lactoferrin Production: A Systematic Review of the Latest Analytical Methods

Background: Lactoferrin is a major functional protein involved in maintaining human health, which possesses antioxidant, anti-inflammatory, antibacterial, and antiviral properties. Therefore, it can be used to support the treatment of viral and bacterial diseases, as well as in cancer prevention. Lactoferrin-manufacturing processes may compromise its protein structure and function, so it is necessary to establish reliable analytical methods for production efficiency and quality control purposes. This paper reviews the lactoferrin production processes, summarising the methods using various matrices (milk, milk powder, infant formula, whey, bovine lactoferrin lyophilised powder, yoghurt, colostrum, and human milk), the most popular purification methods, and sample preparation. Material and methods: The Medline and Embase databases were searched using the following phrases: ”lactoferrin” and “purification” or “isolation” or “extraction” or “separation”. The search was limited to recent studies from the last five years published in English up until 12 March 2025. Of the 573 articles identified, 17 were reviewed. Results: Lactoferrin purification and determination methods depend on the matrix used. The latest research focuses on improving parameters of lactoferrin determination, shortening time, improving efficiency or limiting costs, and even reducing toxicity by changing the reagents. The method of separating lactoferrin using magnetic beads or nanoparticles has been developed, as well as the determination parameters using high-performance liquid chromatography (HPLC). Conclusions: The current lactoferrin production techniques are characterised by increased efficiency and quality, but they require standardisation of the purification process depending on the matrix. The latest Lf determination methods are highly precise, and most of them produce high-quality Lf. This allows to introduce on the market a higher quality product, which can significantly improve standard approaches.