Review Article Cites HemoVoid™ & HemogloBind™ for RBC Proteome Analysis
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Biotech Support Group reports on a recent review article which describes the simplicity and efficiency of their proteomic sample preparation technology for selectively depleting hemoglobin, to help solve the dynamic range problem for comprehensive erythrocyte proteome analysis.
The citation is: Barasa, Benjamin, and Monique Slijper. "Challenges for red blood cell biomarker discovery through proteomics." Biochimica et Biophysica Acta (BBA)-Proteins and Proteomics 1844.5 (2014): 1003-1010.
In brief, this review describes the many challenges to generate in-depth RBC proteome analysis, such as to obtain pure red blood cells, and to acquire an in-depth proteome, despite the dynamic range problem due to a few highly over-represented RBC proteins - especially hemoglobin which accounts for approximately 97% of the cytosolic mass.
The article states "Hemoglobin can also be depleted from an RBC lysate by employing Hemoglobind- or HemoVoid affinity systems. Hemoglobind consist of an elastomeric poly-electrolytic surface that has been optimized to bind Hb from serum samples with high affinity, and can as well be used to remove Hb from RBC lysates. Walpurgis et al. used a complex matrix to deplete the RBC sample for Hb, named HemoVoid, which is made of a library of different ligand combinations, consisting of several kinds of ionic, aromatic, and polymer ligands. Low abundance proteins in the RBC lysate are captured and enriched by the HemoVoid ligand library, while the high abundance proteins such as Hb and CA-I are thought to quickly saturate the system, and they primarily end up in the flow-through. The high abundance proteins in an RBC lysate can thus be easily separated from the low abundance protein fraction. The chosen Hb-depletion approaches by both Alvarez-Llamas et al. and Walpurgis et al. are well compatible with analysis of the RBC protein fractions by 1D or 2D gel electrophoresis, followed by protein identification through mass spectrometry."
"It is worthwhile to note that the authors describe both our strategies for hemoglobin depletion, as the correct choice will vary with the application. With our own experience and with users such as those referenced in this article, we have gained the necessary knowledge to guide our users to the best option for hemoglobin depletion and/or low abundance enrichment" states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.
