Abstract
Background: Exosomes open exciting new opportunities for advanced drug transport and targeted release. Furthermore, exosomes may be used for vaccination, immunosuppression or wound healing. To fully utilize their potential as drug carriers or immune-modulatory agents, the optimal purity of exosome preparations is of crucial importance.
Methods: Articles describing the isolation and purification of exosomes were retrieved from the PubMed database. Results: Exosomes are often separated from biological fluids containing high concentrations of proteins, lipids and other molecules that keep vesicle purification challenging. A great number of purification protocols have been published, however, their outcome is difficult to compare because the assessment of purity has not been standardized. In this review, we first give an overview of the generation and composition of exosomes, as well as their multifaceted biological functions that stimulated various medical applications. Finally, we describe various methods that have been used to purify small vesicles and to assess the purity of exosome preparations and critically compare the quality of these evaluation protocols. Conclusion: Combinations of various techniques have to be applied to reach the required purity and quality control of exosome preparations.Keywords: Exosomes, extracellular vesicles, ultracentrifugation, proteomics, drug carrier, theranostics, diagnosis.
[http://dx.doi.org/10.2478/rnan-2013-0002]
[http://dx.doi.org/10.3402/jev.v4.26316] [PMID: 25899407]
[http://dx.doi.org/10.1083/jcb.201211138] [PMID: 23420871]
[http://dx.doi.org/10.1146/annurev.micro.091208.073413] [PMID: 20825345]
[http://dx.doi.org/10.1038/nrmicro3525] [PMID: 26373371]
[http://dx.doi.org/10.1128/AEM.03657-12] [PMID: 23315742]
[http://dx.doi.org/10.1038/nrd3978] [PMID: 23584393]
[http://dx.doi.org/10.1128/MMBR.00031-09] [PMID: 20197500]
[http://dx.doi.org/10.1146/annurev-cellbio-101512-122326] [PMID: 25288114]
[http://dx.doi.org/10.1007/978-1-4939-3804-9_18] [PMID: 27665565]
[http://dx.doi.org/10.1124/pr.112.005983] [PMID: 22722893]
[http://dx.doi.org/10.3390/ijms18061153] [PMID: 28555055]
[http://dx.doi.org/10.1038/ncb2502] [PMID: 22660413]
[http://dx.doi.org/10.1242/jcs.128868] [PMID: 24105262]
[http://dx.doi.org/10.1016/j.cub.2009.09.059] [PMID: 19896381]
[http://dx.doi.org/10.1038/nri2567] [PMID: 19498381]
[http://dx.doi.org/10.1016/j.jprot.2010.06.006] [PMID: 20601276]
[http://dx.doi.org/10.1038/srep22519] [PMID: 26931825]
[http://dx.doi.org/10.1073/pnas.1521230113] [PMID: 26858453]
[http://dx.doi.org/10.1080/20013078.2017.1329476] [PMID: 28717422]
[http://dx.doi.org/10.1083/jcb.101.3.942] [PMID: 2993317]
[PMID: 3597417]
[http://dx.doi.org/10.3389/fimmu.2014.00403] [PMID: 25191326]
[http://dx.doi.org/10.1007/s00018-016-2309-8] [PMID: 27438886]
[http://dx.doi.org/10.1038/ncb1596] [PMID: 17486113]
[http://dx.doi.org/10.1038/sj.leu.2404132] [PMID: 16453000]
[http://dx.doi.org/10.1016/j.gdata.2016.05.010] [PMID: 27330995]
[http://dx.doi.org/10.1038/cr.2014.44] [PMID: 24710597]
[http://dx.doi.org/10.1371/journal.pone.0183915] [PMID: 28850588]
[http://dx.doi.org/10.1093/nar/gks658] [PMID: 22821563]
[http://dx.doi.org/10.1002/wrna.1413] [PMID: 28130830]
[http://dx.doi.org/10.1016/j.canlet.2014.02.004] [PMID: 24513178]
[http://dx.doi.org/10.1371/journal.pone.0003148] [PMID: 18773077]
[http://dx.doi.org/10.1038/nrm2652] [PMID: 19277045]
[http://dx.doi.org/10.1126/science.1153124] [PMID: 18309083]
[http://dx.doi.org/10.1016/j.plipres.2017.03.001] [PMID: 28342835]
[http://dx.doi.org/10.1126/science.1174621] [PMID: 20044567]
[http://dx.doi.org/10.1371/journal.pone.0015353] [PMID: 21179422]
[http://dx.doi.org/10.1186/1476-4598-12-62] [PMID: 23767874]
[http://dx.doi.org/10.1038/ncomms1285] [PMID: 21505438]
[http://dx.doi.org/10.1371/journal.pone.0075348] [PMID: 24205408]
[http://dx.doi.org/10.1111/j.1365-3083.2005.01551.x] [PMID: 15787739]
[http://dx.doi.org/10.1038/sj.ki.5002486] [PMID: 17700640]
[http://dx.doi.org/10.1042/BST0390501] [PMID: 21428928]
[http://dx.doi.org/10.1098/rstb.2013.0502] [PMID: 25135963]
[http://dx.doi.org/10.1186/1471-2164-14-319] [PMID: 23663360]
[http://dx.doi.org/10.1371/journal.pone.0030679] [PMID: 22427800]
[http://dx.doi.org/10.1016/j.febslet.2015.04.036] [PMID: 25937124]
[http://dx.doi.org/10.1038/ncomms3980] [PMID: 24356509]
[http://dx.doi.org/10.1074/jbc.M112.446831] [PMID: 23439645]
[http://dx.doi.org/10.3390/ncrna5010028] [PMID: 30901915]
[http://dx.doi.org/10.1016/j.cell.2019.02.018] [PMID: 30951672]
[http://dx.doi.org/10.1186/s12943-019-0984-4] [PMID: 30849983]
[http://dx.doi.org/10.1016/j.ygyno.2008.04.033] [PMID: 18589210]
[http://dx.doi.org/10.3816/CLC.2009.n.006] [PMID: 19289371]
[http://dx.doi.org/10.1002/cncr.27895] [PMID: 23224754]
[http://dx.doi.org/10.1002/ijc.25376] [PMID: 20473869]
[http://dx.doi.org/10.1038/bjc.2013.8] [PMID: 23361059]
[http://dx.doi.org/10.1152/ajprenal.00148.2013] [PMID: 23946286]
[http://dx.doi.org/10.1161/CIRCGENETICS.110.958975] [PMID: 21642241]
[http://dx.doi.org/10.1183/09031936.00029610] [PMID: 20595154]
[http://dx.doi.org/10.1016/j.jns.2015.12.005] [PMID: 26723991]
[http://dx.doi.org/10.1371/journal.pone.0150154] [PMID: 26930277]
[http://dx.doi.org/10.1073/pnas.0804549105] [PMID: 18663219]
[http://dx.doi.org/10.1373/clinchem.2010.150730] [PMID: 21059829]
[http://dx.doi.org/10.1371/journal.pone.0013247] [PMID: 20949044]
[http://dx.doi.org/10.1155/2015/657086] [PMID: 25695100]
[http://dx.doi.org/10.1074/jbc.M113.452458] [PMID: 23880764]
[http://dx.doi.org/10.1093/jmcb/mjt011] [PMID: 23580760]
[http://dx.doi.org/10.1074/jbc.C113.532267] [PMID: 24398677]
[http://dx.doi.org/10.1016/j.bbrc.2014.07.109] [PMID: 25086355]
[http://dx.doi.org/10.1038/ncomms15287] [PMID: 28508895]
[http://dx.doi.org/10.1073/pnas.1704862114] [PMID: 29073103]
[http://dx.doi.org/10.1016/j.cell.2019.02.029] [PMID: 30951670]
[http://dx.doi.org/10.1074/jbc.M301642200] [PMID: 12639953]
[http://dx.doi.org/10.1111/j.1600-0854.2004.00257.x] [PMID: 15634213]
[http://dx.doi.org/10.1083/jcb.200511072] [PMID: 16785323]
[PMID: 28135905]
[http://dx.doi.org/10.1083/jcb.149.4.901] [PMID: 10811830]
[http://dx.doi.org/10.1038/s41598-019-39882-z] [PMID: 30824759]
[http://dx.doi.org/10.1083/jcb.201002049] [PMID: 20837771]
[http://dx.doi.org/10.1158/0008-5472.CAN-09-2470] [PMID: 20124479]
[http://dx.doi.org/10.1016/j.devcel.2011.08.019] [PMID: 21962903]
[http://dx.doi.org/10.3402/jev.v5.31295] [PMID: 27421995]
[http://dx.doi.org/10.1016/j.bbalip.2012.05.005] [PMID: 22659299]
[http://dx.doi.org/10.1074/jbc.M501112200] [PMID: 15870081]
[http://dx.doi.org/10.1371/journal.pone.0005219] [PMID: 19381331]
[http://dx.doi.org/10.1186/1471-2407-11-108] [PMID: 21439085]
[http://dx.doi.org/10.4161/onci.20897] [PMID: 23170255]
[http://dx.doi.org/10.1038/nature06307] [PMID: 17960135]
[http://dx.doi.org/10.1182/blood-2008-08-174094] [PMID: 19064723]
[http://dx.doi.org/10.1016/j.biocel.2010.10.002] [PMID: 20937409]
[http://dx.doi.org/10.3390/ijms17081296] [PMID: 27517914]
[http://dx.doi.org/10.1146/annurev-cellbio-101512-122422] [PMID: 25000995]
[http://dx.doi.org/10.1371/journal.pbio.1001604] [PMID: 23874151]
[http://dx.doi.org/10.1074/jbc.M114.588046] [PMID: 24951588]
[http://dx.doi.org/10.1074/jbc.M112.445403] [PMID: 23653359]
[http://dx.doi.org/10.1128/JVI.01310-13] [PMID: 23864627]
[http://dx.doi.org/10.1073/pnas.1304266110] [PMID: 24101524]
[http://dx.doi.org/10.1155/2014/619829] [PMID: 24575409]
[http://dx.doi.org/10.3402/jev.v4.27066] [PMID: 25979354]
[http://dx.doi.org/10.1093/intimm/dxh267] [PMID: 15908444]
[http://dx.doi.org/10.1073/pnas.0403453101] [PMID: 15326289]
[http://dx.doi.org/10.1183/09031936.03.00041703] [PMID: 14582906]
[http://dx.doi.org/10.4049/jimmunol.179.3.1969] [PMID: 17641064]
[http://dx.doi.org/10.1097/QAD.0000000000000159] [PMID: 24413309]
[http://dx.doi.org/10.1093/nar/gku347] [PMID: 24838567]
[http://dx.doi.org/10.1002/biof.1407] [PMID: 29341292]
[http://dx.doi.org/10.1007/s00281-010-0233-9] [PMID: 21174094]
[http://dx.doi.org/10.1038/nri3622] [PMID: 24566916]
[http://dx.doi.org/10.1038/ni854] [PMID: 12426563]
[http://dx.doi.org/10.1016/j.bcmd.2005.04.004] [PMID: 15932799]
[http://dx.doi.org/10.3402/jev.v2i0.22492] [PMID: 24223258]
[http://dx.doi.org/10.4049/jimmunol.1203082] [PMID: 23418627]
[http://dx.doi.org/10.1158/0008-5472.CAN-09-3276] [PMID: 20145139]
[http://dx.doi.org/10.1016/j.trsl.2013.10.005] [PMID: 24231336]
[http://dx.doi.org/10.1530/JME-17-0080] [PMID: 28835418]
[http://dx.doi.org/10.1016/j.scr.2013.01.002] [PMID: 23399448]
[http://dx.doi.org/10.1371/journal.pone.0190358] [PMID: 29293592]
[http://dx.doi.org/10.1371/journal.pone.0098667] [PMID: 24905832]
[http://dx.doi.org/10.1111/aji.12311] [PMID: 25164206]
[http://dx.doi.org/10.1186/1758-907X-1-7] [PMID: 20226005]
[http://dx.doi.org/10.7150/ijbs.8.118] [PMID: 22211110]
[http://dx.doi.org/10.1186/1471-2407-12-22] [PMID: 22251860]
[http://dx.doi.org/10.1007/s00018-011-0689-3] [PMID: 21560073]
[http://dx.doi.org/10.1002/art.10312] [PMID: 12115179]
[http://dx.doi.org/10.3389/fonc.2012.00038] [PMID: 22649786]
[http://dx.doi.org/10.1073/pnas.1014194107] [PMID: 21059916]
[http://dx.doi.org/10.1002/pmic.200800793] [PMID: 19415654]
[http://dx.doi.org/10.1016/j.lfs.2011.06.020] [PMID: 21763319]
[http://dx.doi.org/10.1007/s00005-016-0453-3] [PMID: 28101591]
[http://dx.doi.org/10.1038/nature15756] [PMID: 26524530]
[http://dx.doi.org/10.3390/cancers11030326] [PMID: 30866509]
[http://dx.doi.org/10.1158/0008-5472.CAN-10-4455] [PMID: 21478294]
[http://dx.doi.org/10.1038/ncb3169] [PMID: 25985394]
[http://dx.doi.org/10.1038/nature15376] [PMID: 26479035]
[http://dx.doi.org/10.1016/j.cell.2012.11.024] [PMID: 23260141]
[http://dx.doi.org/10.1161/CIRCGENETICS.110.958363] [PMID: 20959591]
[http://dx.doi.org/10.1016/j.semcdb.2015.02.007] [PMID: 25704308]
[http://dx.doi.org/10.1523/JNEUROSCI.5699-09.2010] [PMID: 20484626]
[http://dx.doi.org/10.1083/jcb.201504057] [PMID: 26598621]
[http://dx.doi.org/10.1073/pnas.0603838103] [PMID: 16837572]
[http://dx.doi.org/10.1007/s10571-015-0294-3] [PMID: 26908139]
[http://dx.doi.org/10.1038/srep45034] [PMID: 28338013]
[http://dx.doi.org/10.1007/978-1-4939-8879-2_21] [PMID: 30378059]
[http://dx.doi.org/10.1111/boc.201400095] [PMID: 25923825]
[http://dx.doi.org/10.1186/s12907-015-0005-5] [PMID: 25883534]
[http://dx.doi.org/10.1038/s41571-018-0036-9] [PMID: 29795272]
[http://dx.doi.org/10.1016/j.cca.2018.09.007] [PMID: 30194933]
[http://dx.doi.org/10.1186/1479-5876-3-10] [PMID: 15740633]
[http://dx.doi.org/10.1371/journal.pone.0004942] [PMID: 19319200]
[http://dx.doi.org/10.1371/journal.pone.0003377] [PMID: 18852879]
[http://dx.doi.org/10.1038/nbt.1807] [PMID: 21423189]
[http://dx.doi.org/10.1093/nar/gks463] [PMID: 22618874]
[http://dx.doi.org/10.1038/mt.2012.180] [PMID: 23032975]
[http://dx.doi.org/10.1016/j.jconrel.2015.03.033] [PMID: 25836593]
[PMID: 24747178]
[http://dx.doi.org/10.1016/j.nano.2015.10.012] [PMID: 26586551]
[http://dx.doi.org/10.1016/j.biomaterials.2013.11.083] [PMID: 24345736]
[http://dx.doi.org/10.1038/mt.2010.105] [PMID: 20571541]
[http://dx.doi.org/10.1080/14756366.2017.1292263] [PMID: 28262028]
[http://dx.doi.org/10.1038/srep38541] [PMID: 27941871]
[http://dx.doi.org/10.1002/jcp.27615] [PMID: 30317632]
[http://dx.doi.org/10.1016/j.bbrc.2019.03.119]
[http://dx.doi.org/10.1371/journal.pone.0214545] [PMID: 30925190]
[http://dx.doi.org/10.1080/20013078.2018.1560809] [PMID: 30651940]
[http://dx.doi.org/10.1080/20013078.2018.1535750] [PMID: 30637094]
[http://dx.doi.org/10.3390/v7072810] [PMID: 26205405]
[http://dx.doi.org/10.1186/s12977-014-0102-z] [PMID: 25407601]
[http://dx.doi.org/10.1080/19336918.2016.1251000] [PMID: 27875097]
[http://dx.doi.org/10.1186/1471-2407-12-421] [PMID: 22998595]
[PMID: 29187456]
[http://dx.doi.org/10.1074/jbc.M109.041152] [PMID: 19801663]
[http://dx.doi.org/10.1158/0008-5472.CAN-07-6538] [PMID: 18829542]
[http://dx.doi.org/10.1038/srep16662] [PMID: 26567861]
[http://dx.doi.org/10.1371/journal.pone.0089534] [PMID: 24586853]
[http://dx.doi.org/10.1016/j.bbrc.2016.10.019] [PMID: 27725157]
[http://dx.doi.org/10.1016/j.bbrc.2015.03.172] [PMID: 25849885]
[http://dx.doi.org/10.1021/acs.chemrev.7b00534] [PMID: 29384376]
[http://dx.doi.org/10.1002/0471143030.cb0322s30]
[http://dx.doi.org/10.1016/j.jconrel.2016.02.037] [PMID: 26941033]
[http://dx.doi.org/10.1038/nmeth.4185] [PMID: 28245209]
[http://dx.doi.org/10.13070/mm.en.5.1450]
[http://dx.doi.org/10.1371/journal.pone.0170628] [PMID: 28114422]
[http://dx.doi.org/10.1186/s12967-016-0935-4] [PMID: 27317346]
[http://dx.doi.org/10.1371/journal.pone.0145686] [PMID: 26690353]
[http://dx.doi.org/10.1007/978-1-4939-7652-2_7] [PMID: 29388137]
[http://dx.doi.org/10.1016/j.nano.2015.01.003] [PMID: 25659648]
[http://dx.doi.org/10.3390/cells7120273] [PMID: 30558352]
[http://dx.doi.org/10.1080/20013078.2017.1294340] [PMID: 28386391]
[http://dx.doi.org/10.1186/s12967-017-1374-6] [PMID: 29316942]
[http://dx.doi.org/10.1038/srep33641] [PMID: 27640641]
[http://dx.doi.org/10.1371/journal.pone.0198820] [PMID: 29889903]
[http://dx.doi.org/10.1038/srep23978] [PMID: 27068479]
[http://dx.doi.org/10.1371/journal.pone.0186534] [PMID: 29023592]
[http://dx.doi.org/10.1038/srep14664] [PMID: 26419333]
[http://dx.doi.org/10.1002/biot.201600699] [PMID: 28166394]
[http://dx.doi.org/10.1039/B916199F] [PMID: 20126692]
[http://dx.doi.org/10.1021/acs.analchem.5b01636] [PMID: 26291637]
[http://dx.doi.org/10.1021/pr800225z] [PMID: 18570454]
[http://dx.doi.org/10.1021/acs.analchem.8b05749] [PMID: 30832474]
[http://dx.doi.org/10.1039/C6LC01007E] [PMID: 27722618]
[http://dx.doi.org/10.1063/1.4927494] [PMID: 26339309]
[http://dx.doi.org/10.1039/c2lc21304d] [PMID: 22334376]
[http://dx.doi.org/10.1021/acsnano.7b02277] [PMID: 28679045]
[http://dx.doi.org/10.1186/s12951-018-0433-3] [PMID: 30612562]
[http://dx.doi.org/10.1016/S0022-1759(00)00321-5] [PMID: 11150547]
[http://dx.doi.org/10.1039/C8NR02871K] [PMID: 30014056]
[http://dx.doi.org/10.1016/j.ymeth.2012.01.002] [PMID: 22285593]
[http://dx.doi.org/10.1002/pros.23393] [PMID: 28762517]
[http://dx.doi.org/10.1021/acssensors.9b00060] [PMID: 30915846]
[http://dx.doi.org/10.1111/jth.12602] [PMID: 24818656]
[http://dx.doi.org/10.1111/j.1538-7836.2009.03654.x] [PMID: 19840362]
[http://dx.doi.org/10.1007/s10544-012-9642-y] [PMID: 22391880]
[http://dx.doi.org/10.1007/s11051-008-9435-7]
[http://dx.doi.org/10.1016/j.colsurfb.2011.05.013] [PMID: 21640565]
[http://dx.doi.org/10.3402/jev.v1i0.19179] [PMID: 24009887]
[http://dx.doi.org/10.1186/s12575-019-0092-2] [PMID: 30918474]
[http://dx.doi.org/10.1021/acsami.6b09442] [PMID: 27617891]
[http://dx.doi.org/10.1088/2050-6120/aad57d] [PMID: 30039805]
[http://dx.doi.org/10.1038/srep27290] [PMID: 27264341]
[http://dx.doi.org/10.1016/j.bbamcr.2018.09.008] [PMID: 30290236]
[http://dx.doi.org/10.3402/jev.v4.25530] [PMID: 25833224]
[http://dx.doi.org/10.1002/cyto.a.10115] [PMID: 14750130]
[http://dx.doi.org/10.1161/ATVBAHA.111.244616] [PMID: 22328775]
[http://dx.doi.org/10.1002/cyto.a.22644] [PMID: 25688721]
[PMID: 26484737]
[http://dx.doi.org/10.1080/20013078.2017.1344087] [PMID: 28804597]
[http://dx.doi.org/10.1002/cyto.a.22795] [PMID: 26651033]
[http://dx.doi.org/10.3402/jev.v2i0.19861] [PMID: 24009896]
[http://dx.doi.org/10.1016/j.nano.2011.04.003] [PMID: 21601655]
[http://dx.doi.org/10.1016/j.ymeth.2015.03.028] [PMID: 25843788]
[http://dx.doi.org/10.1155/2016/2639728] [PMID: 27118976]
[http://dx.doi.org/10.1021/la960224o]
[http://dx.doi.org/10.1111/j.1423-0410.2008.01151.x] [PMID: 19175566]
[http://dx.doi.org/10.1016/j.ab.2012.06.004] [PMID: 22691960]
[http://dx.doi.org/10.1021/ac200195n] [PMID: 21434639]
[http://dx.doi.org/10.1002/jcb.21923] [PMID: 18802920]
[http://dx.doi.org/10.3389/fphys.2018.01169] [PMID: 30197601]
[PMID: 28158410]
[http://dx.doi.org/10.3791/58814] [PMID: 30582600]
[http://dx.doi.org/10.1016/j.yexcr.2019.03.014] [PMID: 30857972]
[http://dx.doi.org/10.3389/fphys.2018.01479] [PMID: 30405435]
[PMID: 26261491]
[http://dx.doi.org/10.3402/jev.v4.27031] [PMID: 26194179]
[http://dx.doi.org/10.1038/s41598-019-41800-2] [PMID: 30926864]
[http://dx.doi.org/10.1080/15476286.2016.1270005] [PMID: 28005467]
[http://dx.doi.org/10.1038/s41598-018-38320-w] [PMID: 30755672]
[http://dx.doi.org/10.2147/CMAR.S178326] [PMID: 30464595]
[http://dx.doi.org/10.1080/15476286.2018.1564465] [PMID: 30604646]
[http://dx.doi.org/10.1038/srep33935] [PMID: 27659060]
[http://dx.doi.org/10.1038/srep23550] [PMID: 27009329]
[http://dx.doi.org/10.1371/journal.pone.0205496] [PMID: 30379855]
[http://dx.doi.org/10.1002/pmic.201800257] [PMID: 30411858]
[http://dx.doi.org/10.1016/j.placenta.2016.09.008] [PMID: 27780544]
[http://dx.doi.org/10.1515/cclm-2018-1297] [PMID: 30990781]
[http://dx.doi.org/10.1080/20013078.2018.1442088] [PMID: 29535850]
[http://dx.doi.org/10.1021/acs.analchem.7b04374] [PMID: 29870225]
[http://dx.doi.org/10.1021/ac504861d] [PMID: 25674701]
[http://dx.doi.org/10.3892/ijmm.2017.3080] [PMID: 28737826]
[http://dx.doi.org/10.1080/19336918.2017.1279784] [PMID: 28146372]
[http://dx.doi.org/10.3389/fchem.2017.00102] [PMID: 29209607]
[http://dx.doi.org/10.1074/mcp.M112.021303] [PMID: 23230278]
[http://dx.doi.org/10.1080/14789450.2017.1260450] [PMID: 27838931]
[http://dx.doi.org/10.1016/j.jprot.2013.10.035] [PMID: 24211767]
[http://dx.doi.org/10.1021/pr101060v] [PMID: 21309581]
[http://dx.doi.org/10.3390/proteomes6030032] [PMID: 30200280]
[http://dx.doi.org/10.1007/s00018-018-2773-4] [PMID: 29441425]
[http://dx.doi.org/10.1021/acs.analchem.7b03119] [PMID: 28949504]
[http://dx.doi.org/10.1080/20013078.2018.1490143] [PMID: 29988836]
[http://dx.doi.org/10.1007/978-1-62703-405-0_16] [PMID: 23585095]
[http://dx.doi.org/10.1039/C6MB00082G] [PMID: 27030573]