Rapid fractionation of bovine transferrin using immobilized gangliosides

Bovine transferrin (BTF) was fractionated from bovine whey using gangliosid e affinity chromatography. After loading the immobilized matrix with a 2% w hey solution, the matrix was washed with sodium acetate buffer at pH 4 cont aining 1 M NaCl before elution of BTF with sodium phosphate buffers at pH 7 . Concanavalin-A affinity and ion exchange chromatography were used for fur ther purification. The ganglioside column showed a 74.2% BTF recovery from whey and BTF was enriched to 61% purity with ion exchange chromatography. B ovine transferrin was identified by SDS-PAGE and western analysis. The Conc anavalin-A affinity and ion exchange chromatography steps enriched BTF in t he samples and removed other whey proteins from ganglioside purified fracti ons. These results indicate that immobilized ganglioside can be used to fra ctionate BTF from bovine whey. Our novel ganglioside affinity chromatograph y is rapid and efficient for the fractionation of BTF from whey.