In this study the antibacterial activity of lactoferrin against Cronobacter sakasakii (C. sakasakii) as a foodborne pathogen was investigated. The bovine lactoferrin (bLf) was isolated and purified from bovine colostrum by FPLC chromatography and visualized using (SDS-PAGE). Purification efficiency was 90%. C. sakasakii was isolated from a total of 100 samples of infant formula milk powder (IFMP) samples collected randomly from Qena city groceries and supermarkets using HiCrome Enterobacter sakazakiiagar followed by biochemical testing. C. sakasakii was detected in 21% of the total samples examined. The antibacterial effect of bLF on C. sakasakii isolates was tested via performing disk diffusion method using Mueller–Hinton agar. The results revealed that bLF at concentration of 10mg/ml showed the maximum inhibitory effect whereas the least inhibitory effect was recorded at concentration of 1mg/ml. The results indicated that bLF may be useful for inhibition of C. sakasakii in infant formula through supplementation or fortification. More attention should be paid during manufacture and handling of IFMP.