Cleanascite™ from Biotech Support Group for lipid clarification in RBC proteins and immunological research

MONMOUTH JUNCTION, N.J. — Cleanascite™ removes excess lipids such as micelles with lipidated proteins and serves as a potential, non-denaturing methods for purification of lipidated proteins. Cleanascite™ removes lipids and allows for sample preparation using gel filtration, reversed-phase HPLC, hydrophobic interaction chromatography (HIC) and mass spectrometry.

Now a published paper in the journal of Immunology and Cell Biology titled, “Red blood cells release factors with growth and survival bioactivities for normal and leukemic T cells” by Antunes et al has cited the successful use of Cleanascite ™ for removing lipids within red blood cells. This research opens up new possibilities in the field of studying RBC released factors and how immunological factors such as T cells, B-cells and CD factors work on in vitro cultured human RBC. Proteins are important constituents of the red blood cell plasma membrane. Studying cell growth, survival and proliferation by analyzing the clarified supernatant from the RBC membrane proteins obtained after the ultracentrifugation step allows scientists to learn more about how proteinaceous factors such as EDGSF (for erythrocyte-derived growth and survival factor) function in cells. So what this allows for is the study of primary structure of the various RBC membrane proteins, understanding definition of specific mutations in various RBC phenotypes, and detailed biophysical characterization of membrane properties of normal and mutant RBCs for forming models of the molecular and structural basis for RBC properties.

According to the published article, “ The RBC-sup was centrifuged at 11 000 g before the in vitro bioactivity assays. For lipid depletion, we added Cleanascite ™ to the RBC-sup in a ratio 1:4 and incubated the mixture first in a rotator at room temperature for 10 min, followed by a further incubation at 4 °C for 30 min, following manufacturer’s instructions. Then, the mixture was centrifuged to remove the resin and the RBC-sup collected and concentrated as indicated above before the in vitro bioactivity assays” . Also, the hydrophobicity of membrane proteins such as actinin, lactoferrin, catalase, enolase, carbonic anhydrase, myeloblastin, lipocalin, B-globin etc has hindered their purification. Cleanascite™ selectively removes lipids, cell debris, lipoproteins, and mucinous impurities from biological samples.

Cleanascite ™ may allow for more effective study of the varied roles of RBC antigens including membrane structural integrity, the transport of molecules through the membrane, as receptors for extracellular ligands, adhesion molecules, enzymes, complement components and regulators, and in glycocalyx formation because it has a high binding capacity for lipids with minimal cross-reactivity with proteins. This makes it ideal for purifying antibodies, recombinant proteins, nucleic acids, proteoglycans. Furthermore two-dimensional gel electrophoresis and mass spectrometry should be subsequently used for protein profile changes in red blood cell membranes. Such a strategy for using Cleanascite ™ initially to purify lipids to allow for analysis of cell proteins makes Cleanascite ™ from Biotech Support Group the ideal reagent to processing, purification and analyze RBC’s in vitro.

Article: Red blood cells release factors with growth and survival bioactivities for normal and leukemic T cells

About Biotech Support Group LLC

Biotech Support Group LLC is a leading developer of proteomic and genomic sample preparation and enrichment products. Its principal products include: AlbuVoid™ for albumin depletion, Cleanascite™ for lipid adsorption and clarification, HemogloBind™ & HemoVoid™ for hemoglobin removal, NuGel™ for functional & chemical proteomics, and ProCipitate™ & ProPrep™ for nucleic acid isolation. For more information, go to www.biotechsupportgroup.com.

CONTACTS:

Dr. Swapan Roy & Matthew Kuruc
Biotech Support Group LLC
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