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10.1038/srep10266 http://scihub22266oqcxt.onion/10.1038/srep10266 C4437317!4437317 !25988257     free     free     free Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\25988257 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       Sci+Rep 2015 ; 5 (ä): 10266 Nephropedia Template TP {{tp|p=25988257 |t=2015. Heparin affinity purification of extracellular vesicles |pdf=|usr=}}{{25988257 }} gab.com Text Heparin affinity purification of extracellular vesicles JO: Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=25988257 #FOAvip Extracellular vesicles (EVs) are lipid membrane vesicles released by cells. They carry active biomolecules including DNA, RNA, and protein which can be transferred to recipient cells. Isolation and purification of EVs from culture cell media and biofluids is still a major challenge. The most widely used isolation method is ultracentrifugation (UC) which requires expensive equipment and only partially purifies EVs. Previously we have shown that heparin blocks EV uptake in cells, supporting a direct EV-heparin interaction. Here we show that EVs can be purified from cell culture media and human plasma using ultrafiltration (UF) followed by heparin-affinity beads. UF/heparin-purified EVs from cell culture displayed the EV marker Alix, contained a diverse RNA profile, had lower levels of protein contamination, and were functional at binding to and uptake into cells. RNA yield was similar for EVs isolated by UC. We were able to detect mRNAs in plasma samples with comparable levels to UC samples. In conclusion, we have discovered a simple, scalable, and effective method to purify EVs taking advantage of their heparin affinity. Twit Text FOAVip Heparin affinity purification of extracellular vesicles ????Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=25988257 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=25988257 #FOAvip English Wikipedia <ref>{{Cite pmid|25988257 }}</ref> Heparin affinity purification of extracellular vesicles #MMPMID25988257 Balaj L ; Atai NA ; Chen W ; Mu D ; Tannous BA ; Breakefield XO ; Skog J ; Maguire CA Sci Rep 2015[May]; 5 (ä): 10266 PMID25988257 show ga Extracellular vesicles (EVs) are lipid membrane vesicles released by cells. They carry active biomolecules including DNA, RNA, and protein which can be transferred to recipient cells. Isolation and purification of EVs from culture cell media and biofluids is still a major challenge. The most widely used isolation method is ultracentrifugation (UC) which requires expensive equipment and only partially purifies EVs. Previously we have shown that heparin blocks EV uptake in cells, supporting a direct EV-heparin interaction. Here we show that EVs can be purified from cell culture media and human plasma using ultrafiltration (UF) followed by heparin-affinity beads. UF/heparin-purified EVs from cell culture displayed the EV marker Alix, contained a diverse RNA profile, had lower levels of protein contamination, and were functional at binding to and uptake into cells. RNA yield was similar for EVs isolated by UC. We were able to detect mRNAs in plasma samples with comparable levels to UC samples. In conclusion, we have discovered a simple, scalable, and effective method to purify EVs taking advantage of their heparin affinity. |Cell Line, Tumor [MESH] |Extracellular Vesicles/genetics/*metabolism [MESH] |HEK293 Cells [MESH] |Heparin/chemistry/*metabolism [MESH] |Human Umbilical Vein Endothelial Cells [MESH] |Humans [MESH] |Microspheres [MESH] |Protein Binding/*physiology [MESH] |RNA, Messenger/genetics [MESH]Heparin affinity purification of extracellular vesicles (epmc).pdf DeepDyve Pubget Overpricing