| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 1200685 | Journal of Chromatography A | 2012 | 8 Pages |
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.
► Food-grade purification of whey-protein–dextran glycates using chromatography. ► Successful scale-up of 160-fold and several fold increase in yield. ► Laser light scattering and MALDI-TOF analysis of whey-protein–dextran glycates. ► Purified product contains oligo-glycates as well as mono-glycates. ► Dextran rated as 10 kDa is 5 kDa by static laser light scattering and MALDI-TOF.
