Lactoferrin Production: A Systematic Review of the Latest Analytical Methods
Source abstract
Abstract
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.
Source: crossref · Quality: baseline
Research context
OpenAlex topics: Infant Nutrition and Health · Pediatric Hepatobiliary Diseases and Treatments · Clinical Nutrition and Gastroenterology
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
Department of Bromatology, Poznan University of Medical Science, Rokietnicka 3 Street, 60-806 Poznan, Poland
Doctoral School, Poznan University of Medical Sciences, Bukowska 70 Street, 60-812 Poznan, Poland
References (100+)
Show source-provided references
1. Kowalczyk, P., Kaczyńska, K., Kleczkowska, P., Bukowska-Ośko, I., Kramkowski, K., and Sulejczak, D. (2022). The Lactoferrin Phenomenon—A Miracle Molecule. Molecules, 27.10.3390/molecules27092941 ↗
2. Lactoferrin in Milk from Different Species10.1016/0305-0491(71)90258-6 ↗
3. The Proteins in Whey
4. Isolation of an Iron-Containing Red Protein from Human Milk10.3891/acta.chem.scand.14-0510 ↗
5. Structure of Human Lactoferrin: Crystallographic Structure Analysis and Refinement at 2·8 Å Resolution10.1016/0022-2836(89)90602-5 ↗
6. Structural Aspects of Human Lactoferrin in the Iron-Binding Process Studied by Molecular Dynamics and Small-Angle Neutron Scattering10.1140/epje/i2018-11720-x ↗
7. Comparative Study of the Primary Structures of Sero-, Lacto- and Ovotransferrin Glycans from Different Species10.1016/0300-9084(88)90283-0 ↗
8. Recent Advances and Prospects in Purification and Heterologous Expression of Lactoferrin10.1002/fbe2.12003 ↗
9. Yang, Z., Jiang, R., Chen, Q., Wang, J., Duan, Y., Pang, X., Jiang, S., Bi, Y., Zhang, H., and Lönnerdal, B. (2018). Concentration of Lactoferrin in Human Milk and Its Variation during Lactation in Different Chinese Populations. Nutrients, 10.10.3390/nu10091235 ↗
10. Effects of Bovine Lactoferrin on the Immature Porcine Intestine10.1017/S0007114513002456 ↗
11. Therapeutic Applications of Whey Protein
12. Lactoferrin Concentrations in Goat Milk throughout Lactation10.1016/j.smallrumres.2008.07.027 ↗
13. Lactoferrin and Immunoglobulin Contents in Camel’s Milk (Camelus bactrianus, Camelus dromedarius, and Hybrids) from Kazakhstan10.3168/jds.S0022-0302(07)72606-1 ↗
14. Lactoferrin Concentration in Buffalo Milk10.4081/ijas.2013.e23 ↗
15. Lactoferrin in Infant Formulas: Effect on Oxidation10.1021/jf0002490 ↗
16. Antiadenovirus Activity of Milk Proteins: Lactoferrin Prevents Viral Infection10.1016/S0166-3542(01)00197-8 ↗
17. Bactericidal Activity of Human Lactoferrin: Sensitivity of a Variety of Microorganisms10.1128/iai.28.3.893-898.1980 ↗
18. The Effects of Recombinant Human Lactoferrin on Immune Activation and the Intestinal Microbiome Among Persons Living with Human Immunodeficiency Virus and Receiving Antiretroviral Therapy10.1093/infdis/jiz042 ↗
19. Lepanto, M.S., Rosa, L., Cutone, A., Conte, M.P., Paesano, R., and Valenti, P. (2018). Efficacy of Lactoferrin Oral Administration in the Treatment of Anemia and Anemia of Inflammation in Pregnant and Non-Pregnant Women: An Interventional Study. Front. Immunol., 9.10.3389/fimmu.2018.02123 ↗
20. Efficacy and Tolerability of Oral Bovine Lactoferrin Compared to Ferrous Sulfate in Pregnant Women with Iron Deficiency Anemia: A Prospective Controlled Randomized Study10.1080/00016340903117994 ↗
21. Oral Administration of Lactoferrin Increases Hemoglobin and Total Serum Iron in Pregnant womenThis Paper Is One of a Selection of Papers Published in This Special Issue, Entitled 7th International Conference on Lactoferrin: Structure, Function, and Applications, and Has Undergone the Journal’s Usual Peer Review Process10.1139/o06-040 ↗
22. Safety and Efficacy of Lactoferrin versus Ferrous Sulphate in Curing Iron Deficiency and Iron Deficiency Anaemia in Hereditary Thrombophilia Pregnant Women: An Interventional Study10.1007/s10534-014-9723-x ↗
23. Lactoferrin Efficacy versus Ferrous Sulfate in Curing Iron Disorders in Pregnant and Non-Pregnant Women10.1177/039463201002300220 ↗
24. RETRACTED ARTICLE: Lactoferrin versus Ferrous Sulphate for the Treatment of Iron Deficiency Anemia during Pregnancy: A Randomized Clinical Trial10.3109/14767058.2015.1049149 ↗
25. Lactoferrin Efficacy versus Ferrous Sulfate in Treatment of Children with Iron Deficiency Anemia10.1055/s-0041-1731073 ↗
26. Comparing Oral Iron Bisglycinate Chelate, Lactoferrin, Lactoferrin with Iron and Iron Polymaltose Complex in the Treatment of Children with Iron Deficiency Anemia10.1016/j.clnesp.2021.08.040 ↗
27. Iron Metabolism in Infants: Influence of Bovine Lactoferrin from Iron-Fortified Formula10.1016/j.nut.2014.07.006 ↗
28. Effect of Lactoferrin Supplementation on Iron Deficiency Anemia in Primary School Children
29. Lactoferrin versus Iron Hydroxide Polymaltose Complex for the Treatment of Iron Deficiency Anemia in Children with Cerebral Palsy: A Randomized Controlled Trial10.1007/s00431-021-04125-9 ↗
30. Lactoferrin for Iron-Deficiency Anemia in Children with Inflammatory Bowel Disease: A Clinical Trial10.1038/s41390-022-02136-2 ↗
31. Ferrous Sulphate Alone Versus Combination of Ferrous Sulphate and Lactoferrin for The Treatment of Iron Deficiency Anemia during Pregnancy and Their Effect on Neonatal Iron Store: A Randomized Clinical Trial10.21608/ejhm.2021.178616 ↗
32. Efficacy and Safety of Oral Lactoferrin Supplementation in Combination with rHuEPO-β for the Treatment of Anemia in Advanced Cancer Patients Undergoing Chemotherapy: Open-Label, Randomized Controlled Study10.1634/theoncologist.2010-0020 ↗
33. Lactoferrin: A Promising New Player in Treatment of Iron Deficiency Anemia in Patients on Regular Hemodialysis: A Randomized Controlled Trial10.4103/1319-2442.393996 ↗
34. Evaluation of Iron Deficiency as a Nutritional Adaptation to Infectious Disease: An Evolutionary Medicine Perspective10.1002/ajhb.20839 ↗
35. Lang, J., Yang, N., Deng, J., Liu, K., Yang, P., Zhang, G., and Jiang, C. (2011). Inhibition of SARS Pseudovirus Cell Entry by Lactoferrin Binding to Heparan Sulfate Proteoglycans. PLoS ONE, 6.10.1371/journal.pone.0023710 ↗
36. The Clinical Efficacy of a Bovine Lactoferrin/Whey Protein Ig-Rich Fraction (Lf/IgF) for the Common Cold: A Double Blind Randomized Study10.1016/j.ctim.2012.12.006 ↗
37. A Double-Blind, Placebo-Controlled, Pilot Study of Bovine Lactoferrin Supplementation in Bottle-fed Infants10.1097/01.mpg.0000243435.54958.68 ↗
38. Efficacy of Bovine Lactoferrin Supplementation in Preventing Late-Onset Sepsis in Low Birth Weight Neonates: A Randomized Placebo-Controlled Clinical Trial10.1093/tropej/fmv044 ↗
39. Effects of Lactoferrin on Infectious Diseases in Japanese Summer: A Randomized, Double-Blinded, Placebo-Controlled Trial10.1016/j.jmii.2020.02.010 ↗
40. Bovine Lactoferrin Supplementation for Prevention of Late-Onset Sepsis in Very Low-Birth-Weight Neonates: A Randomized Trial10.1001/jama.2009.1403 ↗
41. Oral Lactoferrin to Prevent Nosocomial Sepsis and Necrotizing Enterocolitis of Premature Neonates and Effect on T-Regulatory Cells10.1055/s-0034-1371704 ↗
42. Effect of Lactoferrin in Patients with Chronic Hepatitis C: Combination Therapy with Interferon and Ribavirin10.1111/j.1440-1746.2007.04858.x ↗
43. Lactoferrin Inhibits Hepatitis B Virus Infection in Cultured Human Hepatocytes10.1016/S1386-6346(02)00088-8 ↗
44. Inhibition of HBV Infection by Bovine Lactoferrin and Iron-, Zinc-Saturated Lactoferrin10.1007/s00430-008-0100-7 ↗
45. Lactoferrin Inhibits Human Papillomavirus Binding and Uptake in Vitro10.1016/j.antiviral.2004.05.005 ↗
46. The Anti-Papillomavirus Activity of Human and Bovine Lactoferricin10.1016/j.antiviral.2007.03.012 ↗
47. Lactoferrin Prevents Dendritic Cell-Mediated Human Immunodeficiency Virus Type 1 Transmission by Blocking the DC-SIGN—Gp120 Interaction10.1128/JVI.79.5.3009-3015.2005 ↗
48. Bovine Lactoferrampin, Human Lactoferricin, and Lactoferrin 1-11 Inhibit Nuclear Translocation of HIV Integrase10.1007/s12010-016-2059-y ↗
49. Triple Drug Combination of Zidovudine, Efavirenz and Lamivudine Loaded Lactoferrin Nanoparticles: An Effective Nano First-Line Regimen for HIV Therapy10.1007/s11095-016-2048-4 ↗
50. Senapathi, J., Bommakanti, A., Mallepalli, S., Mukhopadhyay, S., and Kondapi, A.K. (2020). Sulfonate Modified Lactoferrin Nanoparticles as Drug Carriers with Dual Activity against HIV-1. Colloids Surf. B Biointerfaces, 191.10.1016/j.colsurfb.2020.110979 ↗
51. Rosa, L., Tripepi, G., Naldi, E., Aimati, M., Santangeli, S., Venditto, F., Caldarelli, M., and Valenti, P. (2021). Ambulatory COVID-19 Patients Treated with Lactoferrin as a Supplementary Antiviral Agent: A Preliminary Study. JCM, 10.10.3390/jcm10184276 ↗
52. Campione, E., Lanna, C., Cosio, T., Rosa, L., Conte, M.P., Iacovelli, F., Romeo, A., Falconi, M., Del Vecchio, C., and Franchin, E. (2021). Lactoferrin as Antiviral Treatment in COVID-19 Management: Preliminary Evidence. Int. J. Environ. Res. Public Health, 18.10.3390/ijerph182010985 ↗
53. Liposomal Lactoferrin as Potential Preventative and Cure for COVID-19
54. Loss of Microbicidal Activity and Increased Formation of Biofilm Due to Decreased Lactoferrin Activity in Patients with Cystic Fibrosis10.1086/423821 ↗
55. Inv-Mediated Apoptosis of Epithelial Cells Infected with Enteropathogenic Yersinia: A Protective Effect of Lactoferrin10.1016/j.resmic.2005.02.007 ↗
56. Both Lactoferrin and Iron Influence Aggregation and Biofilm Formation in Streptococcus Mutans10.1023/B:BIOM.0000027704.53859.d3 ↗
57. Killing of Gram-Negative Bacteria by Lactoferrin and Lysozyme10.1172/JCI115407 ↗
58. Li, C., Sun, Y., He, T., Lu, Y., Szeto, I.M.-Y., Duan, S., Zhang, Y., Liu, B., Zhang, Y., and Zhang, W. (2023). Synergistic Effect of Lactoferrin and Osteopontin on Intestinal Barrier Injury. Int. J. Biol. Macromol., 253.10.1016/j.ijbiomac.2023.127416 ↗
59. Effect of Lactoferrin on Enteric Pathogens10.1016/j.biochi.2008.04.006 ↗
60. Treatment of Lactoferrin and Antimicrobial Peptide N6 on Bacterial Enteritis Caused by Escherichia Coli in Mice10.1139/bcb-2024-0103 ↗
61. Zhang, J., Xin, H., Wang, W., Li, Y., Wu, R., Wei, L., Su, S., Wang, X., Wang, X., and Wang, X. (2024). Investigating the Modulatory Effects of Lactoferrin on Depressed Rats through 16S rDNA Gene Sequencing and LC–MS Metabolomics Analysis. Sci. Rep., 14.10.1038/s41598-024-72793-2 ↗
62. Bovine Lactoferrin and Chimera Lactoferrin Prevent and Destroy Salmonella Typhimurium Biofilms in Caco-2 Cells10.1139/bcb-2024-0100 ↗
63. Inhibition of 7,12-Dimethylbenz[a]Anthracene-Induced Mammary Tumors and DNA Adducts by Garlic Powder10.1093/carcin/13.10.1847 ↗
64. Risiglione, P., Leggio, L., Cubisino, S.A.M., Reina, S., Paternò, G., Marchetti, B., Magrì, A., Iraci, N., and Messina, A. (2020). High-Resolution Respirometry Reveals MPP+ Mitochondrial Toxicity Mechanism in a Cellular Model of Parkinson’s Disease. Int. J. Mol. Sci., 21.10.3390/ijms21217809 ↗
65. A Pilot Study on the Effect of Lactoferrin on Alzheimer’s Disease Pathological Sequelae: Impact of the p-Akt/PTEN Pathway10.1016/j.biopha.2018.12.118 ↗
66. Wang, J., Bi, M., Liu, H., Song, N., and Xie, J. (2015). The Protective Effect of Lactoferrin on Ventral Mesencephalon Neurons against MPP+ Is Not Connected with Its Iron Binding Ability. Sci. Rep., 5.10.1038/srep10729 ↗
67. Lactoferrin from Bovine Colostrum Regulates Prolyl Hydroxylase 2 Activity and Prevents Prion Protein-Mediated Neuronal Cell Damage via Cellular Prion Protein10.1016/j.neuroscience.2014.05.030 ↗
68. Li, Y.-Q., and Guo, C. (2021). A Review on Lactoferrin and Central Nervous System Diseases. Cells, 10.10.3390/cells10071810 ↗
69. Sanches, E., Van De Looij, Y., Sow, S., Toulotte, A., Da Silva, A., Modernell, L., and Sizonenko, S. (2021). Dose-Dependent Neuroprotective Effects of Bovine Lactoferrin Following Neonatal Hypoxia–Ischemia in the Immature Rat Brain. Nutrients, 13.10.3390/nu13113880 ↗
70. Effect of Bovine Lactoferrin as a Novel Therapeutic Agent in a Rat Model of Sepsis-Induced Acute Lung Injury10.1186/s13568-019-0900-8 ↗
71. Li, Y., Li, J., Dong, Y., Wang, C., and Cai, Z. (2024). Bovine Lactoferrin Inhibits Inflammatory Response and Apoptosis in Lipopolysaccharide-Induced Acute Lung Injury by Targeting the PPAR-γ Pathway. Mol. Biol. Rep., 51.10.1007/s11033-024-09436-2 ↗
72. Dose-Dependent Renoprotective Impact of Lactoferrin against Glycerol-Induced Rhabdomyolysis and Acute Kidney Injury10.1016/j.lfs.2022.120646 ↗
73. Lactoferrin Alleviates Gentamicin-Induced Acute Kidney Injury in Rats by Suppressing Ferroptosis: Highlight on ACSL4, SLC7A11, NCOA4, FSP1 Pathways and miR-378a-3p, LINC00618 Expression10.1016/j.fct.2024.115027 ↗
74. Zhang, K., Ma, X., Teng, D., Mao, R., Yang, N., Hao, Y., and Wang, J. (2024). Three in One with Dual-Functional Hydrogel of Lactoferrin/NZ2114/LMSH Promoting Staphylococcus Aureus-Infected Wound Healing. Antibiotics, 13.10.3390/antibiotics13090889 ↗
75. The Preparation of Lactoferrin/Magnesium Silicate Lithium Injectable Hydrogel and Application in Promoting Wound Healing10.1016/j.ijbiomac.2022.09.126 ↗
76. Human Lactoferrin Stimulates Skin Keratinocyte Function and Wound Re-Epithelialization: hLF Stimulates Wound Re-Epithelialization10.1111/j.1365-2133.2010.09748.x ↗
77. Home Treatment Protocol for Dentin Hypersensitivity with Hydroxyapatite-Based Biodynamic Toothpaste and Lactoferrin: A Pilot Study10.23736/S2724-6329.24.05073-3 ↗
78. Yamada, S., Chea, C., Furusho, H., Oda, K., Shiba, F., Tanimoto, K., Tate, S., Miyauchi, M., and Takata, T. (2024). Effects of Novel Lactoferrin Peptides on LPS -induced Alveolar Bone Destruction in a Rat Model. Chem. Biol. Drug Des., 104.10.1111/cbdd.14574 ↗
79. Bovine Lactoferrin and Lactoferricin Derived from Milk: Production and Applications10.1139/o01-230 ↗
80. Lactoferrin Research, Technology and Applications10.1016/j.idairyj.2006.06.013 ↗
81. Current Practices with Commercial Scale Bovine Lactoferrin Production and Alternative Approaches10.1016/j.idairyj.2021.105263 ↗
82. Hancocks, N. (2025, February 15). Lactoferrin Market Set to Surpass €265 Million by 2027. Available online: https://www.nutraingredients.com/Article/2021/04/09/Lactoferrin-market-set-to-surpass-265-million-by-2027/.
83. (2025, March 18). Dairy Reporter 2020. Available online: https://www.dairyreporter.com/Article/2020/10/29/Morinaga-Milk-expanding-lactoferrin-production-capacity-through-German-subsidiary/.
84. Recombinant Human Lactoferrin Expressed in Glycoengineered Pichia Pastoris: Effect of Terminal N-Acetylneuraminic Acid on in Vitro Secondary Humoral Immune Response10.1007/s10719-008-9123-y ↗
85. Yang, P., Wang, J., Gong, G., Sun, X., Zhang, R., Du, Z., Liu, Y., Li, R., Ding, F., and Tang, B. (2008). Cattle Mammary Bioreactor Generated by a Novel Procedure of Transgenic Cloning for Large-Scale Production of Functional Human Lactoferrin. PLoS ONE, 3.10.1371/journal.pone.0003453 ↗
86. Production of Human Lactoferrin in Transgenic Cell Suspension Cultures of Sweet Potato10.1007/s10535-005-0087-5 ↗
87. Expression of Recombinant Human Lactoferrin in Transgenic Alfalfa Plants10.1007/s10535-013-0305-5 ↗
88. Detection and Quantification of Lactoferrin: Innovations, Applications, and Challenges10.1016/j.foodchem.2024.142204 ↗
89. Recombinant Human Lactoferrin: A Valuable Protein for Pharmaceutical Products and Functional Foods10.1016/j.biotechadv.2010.07.002 ↗
90. Expression, Characterization, and Purification of Recombinant Porcine Lactoferrin in Pichia Pastoris10.1006/prep.2001.1607 ↗
91. Yen, C.-C., Wu, P.-Y., Ou-Yang, H., Chen, H.-L., Chong, K.-Y., Chang, R.-L., and Chen, C.-M. (2024). Production of Bioactive Porcine Lactoferrin through a Novel Glucose-Inducible Expression System in Pichia Pastoris: Unveiling Antimicrobial and Anticancer Functionalities. Int. J. Mol. Sci., 25.10.3390/ijms25031818 ↗
92. Lactoferrin-Derived Resistance against Plant Pathogens in Transgenic Plants10.1021/jf400756t ↗
93. Hu, C., Shen, W., Xia, Y., Yang, H., and Chen, X. (2024). Lactoferrin: Current Situation and Future Prospects. Food Biosci., 62.10.1016/j.fbio.2024.105183 ↗
94. Quality Control of Commercial Bovine Lactoferrin10.1007/s10534-018-0098-2 ↗
95. (2025, April 12). Available online: https://www.fda.gov/media/124472/download.
96. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) (2012). Scientific Opinion on Bovine Lactoferrin. EFSA J., 10, 2701.10.2903/j.efsa.2012.2701 ↗
97. Boron-Doped Titania for Separation and Purification of Lactoferrin in Dairy Products10.1016/j.jchromb.2022.123501 ↗
98. Analysis of Bovine Lactoferrin in Infant Formula and Adult Nutritional Products by Optical Biosensor Immunoassay: First Action 2021.0710.1093/jaoacint/qsac066 ↗
99. Bovine Lactoferrin Quantification in Dairy Products by a Simple Immunoaffinity Magnetic Purification Method Coupled with High-Performance Liquid Chromatography with Fluorescence Detection10.1021/acs.jafc.9b06043 ↗
100. An Aptamer Affinity Column for Purification and Enrichment of Lactoferrin in Milk10.1016/j.jchromb.2021.122724 ↗