Determination of native and denatured milk proteins by high-performance size exclusion chromatography.
High-performance size exclusion chromatography on a TSK 3000 SW column was used to separate and quantify the individual milk proteins in their native and/or denatured state. Quantitation of the individual native whey proteins within +/- 10% was achieved. The denatured proteins were dissociated into their monomer units by using sodium dodecyl sulphate and beta-mercaptoethanol. A good separation of the whey protein monomers was achieved and the quantitation of the individual whey proteins within +/- 10% was possible. Under these conditions, alpha s1- and beta-casein were eluted as one peak, whereas kappa-casein and gamma-casein were eluted in the same position as beta-lactoglobulin and alpha-lactalbumin respectively. A method for calculating the areas and hence the concentration of the individual proteins in a mixture of casein and whey proteins is described.
D000818:Animals; D002364:Caseins / Q000032:analysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D007768:Lactalbumin / Q000032:analysis; D007782:Lactoglobulins / Q000032:analysis; D007785:Lactose / Q000032:analysis; D008894:Milk Proteins / Q000032:analysis*; D011489:Protein Denaturation; D012710:Serum Albumin, Bovine / Q000032:analysis; D005719:gamma-Globulins / Q000032:analysis
D002364:Caseins; D007782:Lactoglobulins; D008894:Milk Proteins; D005719:gamma-Globulins; D012710:Serum Albumin, Bovine; D007768:Lactalbumin; D007785:Lactose