title: Chromatographic purification and characterization of whey protein-dextran glycation products.
abstract: A process for the food-grade preparative-scale production and chromatographic purification of whey protein isolate (WPI)-dextran glycates was developed in this work. The Maillard reaction was used to produce the glycates in aqueous solution. A 5 mL cation exchange column and salt step gradients were utilized to elute the glycated protein at low salt and unreacted protein at high salt. The process was scaled-up 160-fold to an 800 mL column. Glycated products were analyzed by SDS-PAGE, BCA protein assay and glycoprotein carbohydrate estimation kit, MALDI-TOF and static and dynamic light scattering. Glycated protein was relatively pure, containing only traces of beta-lactoglobulin, and it was heterogeneous due to oligo-glycation. It had a molecular mass of 26-35 kDa by static light scattering, and 22-67 kDa by MALDI-TOF. Mono-glycated protein would have been 23.8 kDa from beta-lactoglobulin (18.6 kDa) and dextran (5.2 kDa). This work demonstrated the utility of cation exchange chromatography for the large-scale purification of glycated proteins using food-grade chemicals and procedures.
authors.affiliation: Department of Food Science, University of Wisconsin, 1605 Linden Drive, Madison, WI 53706, USA.
authors.forename: Tejashree
authors.initials: T
authors.lastname: Bund
authors.forename: Susann
authors.initials: S
authors.lastname: Allelein
authors.forename: Abhiram
authors.initials: A
authors.lastname: Arunkumar
authors.forename: John A
authors.initials: JA
authors.lastname: Lucey
authors.forename: Mark R
authors.initials: MR
authors.lastname: Etzel
mesh_terms: D000818:Animals; D002412:Cations / Q000737:chemistry; D002417:Cattle; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003911:Dextrans / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D057446:Hydrodynamics; D008027:Light; D015416:Maillard Reaction; D008894:Milk Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010316:Particle Size; D012542:Scattering, Radiation; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D000067816:Whey Proteins
chemical_list: D002412:Cations; D003911:Dextrans; D008894:Milk Proteins; D000067816:Whey Proteins