Oligonucleotide probes and uses thereof
Methods and compositions are provided to identify oligonucleotides that bind targets of interest. The targets include tissues, cells, circulating biomarkers such as microvesicles, including those derived from various diseases. The oligonucleotides can be used in diagnostic and therapeutic applications.
1. A method of enriching an oligonucleotide probe library comprising a plurality of oligonucleotides, the method comprising:
(a) providing a planar support arrayed with a plurality of samples, wherein a portion of the plurality of samples are cancer samples and another portion of the plurality of samples are non-cancer control samples, and wherein each of the plurality of samples comprises microvesicles;
(b) contacting the plurality of samples arrayed on the planar support with the plurality of oligonucleotides; and
(c) recovering members of the oligonucleotide probe library that bound to members of the plurality of samples, thereby enriching the oligonucleotide probe library.
2. The method of claim 1 , further comprising repeating steps (a)-(c) at least 5 times.
3. The method of claim 1 , wherein the unenriched oligonucleotide probe library comprises at least 10 6 different oligonucleotide sequences.
4. The method of claim 1 , wherein the plurality of cancer samples comprises a bodily fluid or a fraction or derivative thereof.
5. The method of claim 4 , wherein the bodily fluid comprises at least one of 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, pre-ejaculatory fluid, female ejaculate, sweat, fecal matter, hair oil, tears, cyst fluid, pleural fluid, peritoneal fluid, malignant 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, and other lavage fluids.
6. The method of claim 4 , wherein the bodily fluid comprises blood or a derivative thereof.
7. The method of claim 1 , further comprising isolating the microvesicles prior to step (a).
8. The method of claim 7 , wherein isolating the microvesicles comprises at least one of filtration, ultracentrifugation, affinity isolation, size exclusion chromatography and polymer precipitation.
9. The method of claim 8 , wherein the polymer comprises polyethylene glycol (PEG).
10. The method of claim 1 , wherein the planar support comprises a well, a glass slide or a membrane.
11. The method of claim 10 , wherein the membrane comprises a polymer membrane or a nitrocellulose membrane.
12. The method of claim 1 , wherein the plurality of cancer samples differs from the plurality of control samples according to a phenotype of interest, some or all members of the plurality of cancer samples and plurality of control samples are from different sections of a same substrate, and some or all members of the plurality of cancer samples and plurality of control samples are scraped or microdissected from the same substrate.
13. A method of characterizing a cancer in a sample comprising:
(a) enriching a plurality of oligonucleotides to characterize the cancer according to the method of claim 1 ;
(b) arraying at least one sample on a substrate;
(c) contacting the substrate with the plurality of oligonucleotides enriched in step (a); and
(d) measuring a presence or level of a complex formed between members of the plurality of oligonucleotides and the samples arrayed on the substrate, wherein the presence or level is used to characterize the cancer.
14. The method of claim 13 , wherein the sample comprises microvesicles isolated from a bodily fluid.
15. The method of claim 14 , 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.
16. The method of claim 13 , wherein the measuring comprises sequencing, amplification, hybridization, gel electrophoresis, chromatography, or visualization.
17. The method of claim 16 , wherein the hybridization comprises contacting the sample with at least one labeled probe that is configured to hybridize with members of the plurality of oligonucleotides.
18. The method of claim 16 , wherein the visualization comprises visualizing a signal linked directly or indirectly to members of the plurality of oligonucleotides.