Optimizing red blood cell protein extraction for biomarker quantitation with mass spectrometry
Stephan Klatt, Anne Roberts, Amber Lothian, Colin L Masters, Roberto Cappai, Christopher Fowler, Blaine R Roberts
ANALYTICAL AND BIOANALYTICAL CHEMISTRY | SPRINGER HEIDELBERG | Published : 2020
Red blood cells (RBC) are the most common cell type found in blood. They might serve as reservoir for biomarker research as they are anuclear and lack the ability to synthesize proteins. Not many biomarker assays, however, have been conducted on RBC because of their large dynamic range of proteins, high abundance of lipids, and hemoglobin interferences. Here, we developed a semiquantitative mass spectrometry-based assay that targeted 144 proteins and compared the efficiency of urea, sodium deoxycholate, acetonitrile, and HemoVoid™ in their extraction of the RBC proteome. Our results indicate that protein extraction with HemoVoid™ led to hemoglobin reduction and increased detection of low abu..View full abstract
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Awarded by National Health and Medical Research Council
We acknowledge funding and support; BR receives partial support from the National Health and Medical Research Council (1061550 & 1138673), and Motor Neuron Disease Research Institute of Australia and the Neuroproteomics Facility. BR and SK receive support from the Michael J. Fox Foundation (MJFF, Parkinson's Research). BR receives support from Alzheimer's Disease Drug Discovery Foundation. BR receives research support from Agilent Technologies and e-MSion, Inc.