MONMOUTH JUNCTION, N.J. - April 24, 2021 - Biotech Support Group reports on a recent research article describing the simplicity and efficiency of two depletion technologies – one for Albumin and one Hemoglobin, to quantify proteome abundance changes upon biphasic calcium phosphate (BCP) ceramics embedded in blood clots.
The citation is:
Jing, Lun, et al. "PROTEOMIC ANALYSIS IDENTIFIED LBP AND CD14 AS KEY PROTEINS IN BLOOD/BIPHASIC CALCIUM PHOSPHATE MICROPARTICLE INTERACTIONS (https://www.sciencedirect.com/science/article/pii/S1742706121002282)." Acta Biomaterialia (2021).
Immediately upon implantation, scaffolds for bone repair are exposed to the patient's blood. Biphasic calcium phosphate (BCP) ceramics are considered as the gold standard in bone reconstruction surgery. Here, in a LC-MS/MS proteomic study, the authors compared the differentially expressed blood proteins (plasma and blood cell proteins) and the deregulated signaling pathways of osteogenic and fibrogenic blood composites. The article describes use of HemoVoid™ for depletion of Hemoglobin prior to LC-MS analysis, "each composite material or 4 blood clots were pooled into 3ml of cooled lysis buffer [HEPES 50mM (pH 7.4); NaCl 150mM; EDTA 20mM (pH 8); CHAPS 1%; DTT 1mM; Protease and Phosphatase inhibitor cocktail]. They were let on ice for 30min with regular vortexings and centrifuged at 4°C, 8000g, 15 min. The supernatants were collected, and hemoglobin depleted using several HemoVoid™ columns (Biotech Support Group). When indicated, albumin was also partially removed using AlbuVoid™ depletion reagent kit (Biotech Support Group) following the manufacturer's instructions." From these enrichment steps, the investigators found respectively 80 and 92 proteins differentially expressed between blood clot and BCP 80-200 or BCP 200-500 blood composites. After albumin depletion, analysis of the significant deregulated proteins showed that 27 signaling pathways significantly changed in blood cells stimulated with BCP 80-200 particles compared to blood cells in blood clot, whereas only 6 of these pathways were significantly deregulated with BCP 200-500 particles. These data obtained after low abundance protein enrichment confirmed that the acute phase response protein, as well as LXR and. FXR pathways, were highly modulated in BCP blood composites but, conversely to what was observed without albumin depletion, these 3 pathways were more strongly altered by the BCP 80- 200 particles. Understanding the molecular events following the interactions of the patient's blood with biomaterials is crucial in the development of bone substitutes at least as effective as bone grafting.
"Here is an excellent report demonstrating a very important advantage of the variety of sample prep products that we supply. ", states Swapan Roy, Ph.D., President and Founder of Biotech Support Group.
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