Depletion of highly abundant proteins in blood plasma by hydrophobic interaction chromatography for proteomic analysis.
The proteomic analysis of plasma is extremely complex due to the presence of few highly abundant proteins. These proteins have to be depleted in order to detect low abundance proteins, which are likely to be of biomedical interest. In this work it was investigated the applicability of hydrophobic interaction chromatography (HIC) as a plasma fractionation method prior to two-dimensional gel electrophoresis (2DGE). The average hydrophobicity of the 56 main plasma proteins was calculated. Plasma proteins were classified as low, medium and highly hydrophobic through a cluster analysis. The highly abundant proteins showed a medium hydrophobicity, and therefore a HIC step was designed to deplete them from plasma. HIC performance was assessed by 2DGE, and it was compared to that obtained by a commercial immuno-affinity (IA) column for albumin depletion. Both methods showed similar reproducibility. HIC allowed partially depleting alpha-1-antitrypsin and albumin, and permitted to detect twice the number of spots than IA. Since albumin depletion by HIC was incomplete, it should be further optimized for its use as a complementary or alternative method to IA.
Mahn
Andrea
A
Department of Chemical Engineering, University of Santiago of Chile, Santiago, Chile. amahn_2000@yahoo.es
Reyes
Alejandro
A
Zamorano
Mauricio
M
Cifuentes
Wildo
W
Ismail
Maritza
M
D000818:Animals; D001798:Blood Proteins / Q000737:chemistry / Q000302:isolation & purification*; D002853:Chromatography, Liquid / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional; D057927:Hydrophobic and Hydrophilic Interactions; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000302:isolation & purification
D001798:Blood Proteins; D012709:Serum Albumin; D000515:alpha 1-Antitrypsin