Comparison of capillary electrophoresis with traditional methods to analyse bovine whey proteins.
The separation of the four major whey proteins by sodium dodecyl sulphate (SDS)-capillary gel electrophoresis (CGE) is described. Whilst commercially purified whey proteins could be analysed using the recommended protocol, the more complex nature of an acid whey and a reconstituted whey protein concentrate (WPC) powder necessitated considerable refinement of the CGE sample buffer. Individual whey proteins in the acid whey and WPC samples were then also separated and quantitated using capillary zone electrophoresis, polyacrylamide gel electrophoresis (PAGE) and HPLC methods and the results were compared. The values obtained for alpha-lactalbumin (alpha-Lac) and beta-lactoglobulin (beta-Lg) were consistent throughout the various methods, although size-exclusion HPLC, SDS-PAGE and SDS-CGE could not separate the two beta-Lg variants or the glycosylated form of alpha-Lac from the beta-Lg. There was considerable variation in the values for the bovine serum albumin and immunoglobulin determined by the different methods and it was concluded that none of the methods could satisfactorily quantitate all four whey proteins.
Kinghorn
N M
NM
Food Science Section, New Zealand Dairy Research Institute, Palmerston North.
Norris
C S
CS
Paterson
G R
GR
Otter
D E
DE
D000818:Animals; D002197:Capillary Action; D002417:Cattle; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004586:Electrophoresis / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D007136:Immunoglobulins / Q000302:isolation & purification; D007768:Lactalbumin / Q000302:isolation & purification; D007782:Lactoglobulins / Q000302:isolation & purification; D008894:Milk Proteins / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate; D000067816:Whey Proteins
D007136:Immunoglobulins; D007782:Lactoglobulins; D008894:Milk Proteins; D000067816:Whey Proteins; D012710:Serum Albumin, Bovine; D012967:Sodium Dodecyl Sulfate; D007768:Lactalbumin