Affinity enrichment of bovine lactoferrin in whey.
Bovine lactoferrin was enriched in various whey samples by affinity chromatography using immobilized gangliosides. Bovine gangliosides were isolated from fresh buttermilk using a combination of ultrafiltration and organic extraction. Isolated gangliosides were covalently immobilized onto controlled-pore glass beads. The immobilized matrix contained 66 micrograms of gangliosides per gram of beads. After loading the matrix with reconstituted whey protein isolate (WPI) or whey protein concentrate (WPC), the matrix was washed with sodium phosphate buffer (pH 7) followed by sodium acetate buffer (pH 4) before elution of lactoferrin with 1 M NaCl in sodium acetate buffer. From the intensities of the protein bands in SDS-PAGE, lactoferrin constituted a minimum of 40% of the total protein in the salt eluted sample. WPI, pretrated by heating and ultrafiltration, showed the highest lactoferrin purity among protein sources, while WPI (10% wt/vol) showed the highest recovery. These results show that immobilized gangliosides can be used to enrich the lactoferrin content of whey.
Walsh
M K
MK
Utah State University, Department of Nutrition and Food Sciences, Logan 84322-8700, USA. mkwalsh@cc.usu.edu
Nam
S H
SH
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005732:Gangliosides / Q000737:chemistry / Q000302:isolation & purification; D007781:Lactoferrin / Q000737:chemistry / Q000302:isolation & purification*; D008892:Milk / Q000737:chemistry*; D008894:Milk Proteins / Q000737:chemistry / Q000302:isolation & purification*; D020539:Sequence Analysis, Protein; D000067816:Whey Proteins
D005732:Gangliosides; D008894:Milk Proteins; D000067816:Whey Proteins; D007781:Lactoferrin