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New High-throughput Biomarker Discovery Platform for Exosome Isolation and poly(A)+ mRNA Analysis

Thursday, 12 September 2013 at 12:45

Add to Calendar ▼2013-09-12 12:45:002013-09-12 13:45:00Europe/LondonNew High-throughput Biomarker Discovery Platform for Exosome Isolation and poly(A)+ mRNA AnalysisExosomes and Circulating Biomarkers Summit 2013 in San Diego, CASan Diego, CASELECTBIOenquiries@selectbiosciences.com

We have developed a unique 96-well filterplate which captures exosomes and microvesicles (EMV) from various biological fluids, such as plasma, urine, saliva, cell culture media, etc.  An additional adaptor will accommodate large volumes of culture media and urine (~10-30 mL).    After centrifugation for only 5 min at 2,000 x g, EMV can be trapped on the membrane.  For mRNA analysis, EMV is lysed on the filterplate, and the resultant lysates are transferred to a 96-well oligo(dT)-immobilized microplate for poly(A)+ mRNA isolation followed by cDNA synthesis and PCR.  The results are comparable to those of standard ultracentrifugation method.  In urine, glomeruli-, proximal tubules-, distal tubules-, and collecting ducts-specific mRNAs can be quantified, and are currently used for the analysis of kidney function in areas such as diabetic nephropathy and kidney transplantation.  In plasma, various cell/tissue-specific mRNAs can be quantified, including hematopoietic cells in bone marrow, endothelial cells, neuronal cells in brain, skeletal muscle cells, fat cells, etc.  The identification and quantification of tissue/cell specific mRNAs in EMV are applicable to various biomarker discovery projects.