PROFILING EXTRACELLULAR VESICLES
Methods and compositions are provided for enriching extracellular vesicles, e.g., from bodily fluids. The vesicles may be indicative of various diseases. Vesicle characterization can be used for diagnostic, theranostic and other purposes.
1 . A method of isolating at least one extracellular vesicle from a sample comprising:
(a) contacting the sample with at least one pluronic block copolymer; and
(b) separating extracellular vesicles associated with the pluronic block copolymers from one or more components of the sample thereby isolating the at least one extracellular vesicle.
2 . The method of claim 1 , wherein the at least one pluronic block copolymer comprises F68, F127 or HLB0.80.
3 . The method of claim 1 , wherein the at least one extracellular vesicle comprises vesicles having a diameter between 10 nm and 1000 nm.
4 . The method of claim 1 , wherein the at least one extracellular vesicle comprises vesicles having a diameter between 20 nm and 200 nm.
5 . The method of claim 1 , wherein the at least one extracellular vesicle comprises vesicles having a diameter between 20 nm and 100 nm.
6 . The method of claim 1 , further comprising subjecting the at least one extracellular vesicle to affinity purification, filtration, polymer precipitation, PEG precipitation, ultracentrifugation, a molecular crowding reagent, affinity isolation, affinity selection, or any combination thereof.
7 . The method of claim 1 , wherein the sample comprises a bodily fluid, tissue sample or cell culture.
8 . The method of claim 6 , wherein the bodily fluid comprises peripheral blood, sera, plasma, ascites, urine, cerebrospinal fluid (CSF), sputum, saliva, bone marrow, synovial fluid, aqueous humor, amniotic fluid, cerumen, breast milk, broncheoalveolar lavage fluid, semen, prostatic fluid, cowper's fluid or pre-ejaculatory fluid, female ejaculate, sweat, fecal matter, hair oil, tears, cyst fluid, pleural and peritoneal fluid, pericardial fluid, lymph, chyme, chyle, bile, interstitial fluid, menses, pus, sebum, vomit, vaginal secretions, mucosal secretion, stool water, pancreatic juice, lavage fluids from sinus cavities, bronchopulmonary aspirates, blastocyl cavity fluid, or umbilical cord blood.
9 . (canceled)
10 . The method of claim 1 , further comprising selectively depleting at least one contaminant from the sample prior to step (a), between steps (a) and (b), after step (b), or any combination thereof.
11 . The method of claim 10 , wherein the at least one contaminant comprises at least one highly abundant protein.
12 . The method of claim 1 , further comprising detecting at least one surface antigen associated with the isolated at least one extracellular vesicle.
13 . The method of claim 12 , wherein the at least one surface antigen is selected from Table 3 or Table 6.
14 . The method of claim 1 , further comprising contacting the isolated at least one extracellular vesicle with at least one binding agent, wherein the at least one binding agent comprises a nucleic acid, DNA molecule, RNA molecule, antibody, antibody fragment, aptamer, peptoid, zDNA, peptide nucleic acid (PNA), locked nucleic acid (LNA), lectin, peptide, dendrimer, membrane protein labeling agent, chemical compound, or a combination thereof.
15 . (canceled)
16 . The method of claim 1 , further comprising:
(c) contacting the isolated at least one extracellular vesicle with an aptamer library prior to step (a), between steps (a) and (b), after step (b), or any combination thereof; and
(d) identifying the members of the aptamer library that bound the isolated at least one extracellular vesicle.
17 . The method of claim 16 , wherein the identifying comprises sequencing, hybridization or amplification.
18 . The method of claim 16 , further comprising detecting at least one payload biomarker within the isolated at least one extracellular vesicle.
19 . The method of claim 18 , wherein the one or more payload biomarker comprises at least one nucleic acid, peptide, protein, lipid, antigen, carbohydrate, and/or proteoglycan.
20 . The method of claim 19 , wherein the nucleic acid comprises at least one of DNA, RNA, mRNA, smRNA, microRNA, Y RNA, lincRNA, mitochondrial RNA, snoRNA, snRNA, rRNA, tRNA, siRNA, hnRNA, or shRNA.
21 . (canceled)
22 . (canceled)
23 . The method of claim 16 , wherein the sample is from a subject suspected of having or being predisposed to a disease or disorder.
24 . The method of claim 23 , wherein the disease or disorder comprises a cancer.
25 .- 45 . (canceled)