Hemoglobin depletion from plasma: considerations for proteomic discovery in sickle cell disease and other hemolytic processes.

ABSTRACT:

Purpose

Hemoglobin (Hb) depletion with nickel affinity chromatography has been shown to increase the number of proteins identified in proteomic studies of erythrocytes, but limited data exist on the application of this technique in depletion of Hb from plasma or serum required for clinical biomarker studies. The aim of this study was to explore the potential of using nickel-beads for Hb depletion of plasma.

Experimental design

Nickel–nitrilotriacetic acid (Ni–NTA) affinity chromatography was used to deplete Hb from hemolyzed plasma samples obtained from children with sickle cell disease (SCD, n=7) and normal human plasma (n=4). Ni–NTA-bound proteins were analyzed by one-dimensional GE, followed by in-gel digestion for characterization using an LTQ-Orbitrap hybrid mass spectr

SUBMITTER: Williams LM 

PROVIDER: S-EPMC4364410 | biostudies-literature | 2010 Dec

REPOSITORIES: biostudies-literature