Anion exchange fractionation of serum proteins versus albumin elimination. Anal Biochem 2007 Sep 01;368(1):24-32
Date
07/10/2007Pubmed ID
17618595DOI
10.1016/j.ab.2007.06.004Scopus ID
2-s2.0-34447332788 (requires institutional sign-in at Scopus site) 23 CitationsAbstract
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 (pI values) higher than the pI of albumin, a second fraction containing proteins with pI values in the same range as the pI of albumin, and a third fraction containing proteins with pI values lower than the pI of albumin. This procedure uses common instrumentation, is carried out under denaturing conditions, and takes less than 30min. 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.
Author List
Sahab ZJ, Iczkowski KA, Sang QXMESH terms used to index this publication - Major topics in bold
AlbuminsAnion Exchange Resins
Blood Proteins
Chromatography, Ion Exchange
Humans
Ligands
Male
Models, Biological
Prostate-Specific Antigen
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Triazines
