Anion exchange fractionation of serum proteins versus albumin elimination

Abstract

Elimination of albumin, constituting more than 50% of total serum proteins, allows increased protein loads on immobilized pH gradient (IPG) gels and better visualization of low-abundance proteins; however, it may result in the loss of albumin-bound low-abundance proteins. In this study, we report the prefractionation of serum proteins by batch anion exchange chromatography into three fractions: one containing proteins with isoelectric points (p I values) higher than the p I of albumin, a second fraction containing proteins with p I values in the same range as the p I of albumin, and a third fraction containing proteins with p I values lower than the p I of albumin. This procedure uses common instrumentation, is carried out under denaturing conditions, and takes less than 30 min. We also report the loss of a clinically established prostate cancer serum biomarker, prostate-specific antigen (PSA), after albumin is eliminated using two commercially available albumin elimination kits: one that uses Cibacron Blue F3GA, which achieves albumin depletion through dye–ligand binding, and one that uses specific albumin antibody. The loss of PSA secondary to albumin elimination exceeded that after batch anion exchange serum sample prefractionation.