Purification of lactoferrin isolated from cow's milk by affinity chromatography

Lactoferrin is a multifunctional protein that has various and amazing properties with antimicrobial properties ,can be a good candidate for clinical and commercial applications. Lactoferrin was purified by ion exchange chromatography using CM sephadex C-50 and to confirm the purification, SDS-PAGE tests and the absence of dye in the presence of tetramethylbenzene were used In this study, in order to isolate lactoferrin, first the whey proteins of milk using ion exchange chromatography by Sephadex C-50 was purified and SDSPAGE test was performed to confirm the purification as well as no dye in the presence of tetramethyl benzidine. New Zealand white male rabbits were immunized with 120 µg / ml lactoferrin and using complete and incomplete Freund’s adjuvant for 6 weeks and after serum isolation were confirmed specific antibodies by dot blot and ELISA. To purify of the specific antibody, the precipitation method with 40% ammonium sulphate was used. After confirmation, it was attached to cyanogen-activated Sepharoz. The whey portion of bovine colostrum was passed through the affinity chromatographic column. In order to purify lactoferrin, it reacts to form a complex of antibody antigens, and eventually other unbound compounds are washed and removed. Purification of lactoferrin was confirmed and its concentration was calculated by Bradford test. The results showed that the immunization method was used to produce specific antibodies against Lactoferrin is a suitable method and purification of lactoferrin from bovine milk using affinity chromatography has an efficiency of 64%.