Effect of glycation on the heterogeneity of human serum albumin analysed by reversed-phase high-performance liquid chromatography in a solvent containing formic acid.
Non-enzymic glycation of human serum albumin (HSA) induces a change in its charge heterogeneity that may account for its particular renal clearance in patients with early diabetic nephropathy. A new high-performance liquid chromatographic analysis for the study of HSA heterogeneity is described based on a high content of formic acid in the mobile phase combined with a concave gradient of isopropanol. Under these conditions, native HSA was separated into three individual components (I, II and III). When glycated HSA was analysed, it was found that although the present method is not suitable for the separation of glycated from non-glycated HSA, it shows the effect of glycation in producing changes in HSA heterogeneity that are different from those reported on surface change. This finding suggests an additional factor (probably conformational changes) that is contributing to the heterogeneity of glycated HSA.
Vidal
P
P
Department of Protein Chemistry, Hagedorn Research Laboratory, Gentofte, Denmark.
Nielsen
E
E
Welinder
B S
BS
D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D005561:Formates / Q000737:chemistry*; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D007525:Isoelectric Focusing; D012709:Serum Albumin / Q000378:metabolism*; D012997:Solvents
D005561:Formates; D012709:Serum Albumin; D012997:Solvents; C030544:formic acid; D005947:Glucose