OLIGONUCLEOTIDE PROBES AND USES THEREOF
Methods and compositions are provided for oligonucleotides that bind target biomarkers and allow characterization of a phenotype. The target biomarkers may include microvesicle antigens, including microvesicles derived from various diseases. The characterization may comprise detection, diagnosis, prognosis, theranosis or other characterization of a disease or disorder.
1 - 21 . (canceled)
22 . A method of characterizing a disease or disorder, comprising:
(a) contacting a biological test sample with a plurality of oligonucleotides, wherein the plurality of oligonucleotides comprises at least 20 different oligonucleotide sequences, and wherein each different member of the plurality of oligonucleotides comprises a sequence that is at least 90 percent homologous to any one of SEQ ID NOs. 510599-510763;
(b) detecting a presence or level of complexes formed in step (a) between the plurality of oligonucleotides of and a target in the biological test sample; and
(c) comparing the presence or level detected in step (b) to a reference level from a biological control sample, thereby characterizing the disease or disorder.
23 . The method of claim 22 , wherein the detecting in step (b) comprises performing at least one of sequencing, amplification, and hybridization of the members of the plurality of oligonucleotides within the complexes.
24 . The method of claim 22 , wherein the step of detecting comprises high-throughput sequencing.
25 . The method of claim 22 , wherein the biological test sample and biological control sample each comprise a tissue sample, a cell culture, or a biological fluid.
26 . The method of claim 25 , wherein the biological fluid comprises a bodily fluid.
27 . The method of claim 26 , 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, tears, cyst fluid, pleural fluid, 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.
28 . The method of claim 26 , wherein the bodily fluid comprises blood, serum or plasma.
29 . The method of claim 25 , wherein the biological fluid comprises a microvesicles population.
30 . The method of claim 29 , wherein the complexes are formed between the oligonucleotide or plurality of oligonucleotides and the microvesicles population.
31 . The method of claim 22 , wherein the microvesicle population is isolated using at least one of chromatography, filtration, ultrafiltration, centrifugation, ultracentrifugation, flow cytometry, affinity capture, polymer precipitation, and microfluidics.
32 . The method of claim 22 , wherein at least some members of the plurality of oligonucleotides binds a polypeptide or fragment thereof.
33 . (canceled)
34 . (canceled)
35 . The method of claim 22 , wherein at least some members of the plurality of oligonucleotides binds a microvesicle surface antigen.
36 . The method of claim 22 , wherein the disease or disorder comprises Breast Cancer, Alzheimer's disease, bronchial asthma, Transitional cell carcinoma of the bladder, Giant cellular osteoblastoclastoma, Brain Tumor, Colorectal adenocarcinoma, Chronic obstructive pulmonary disease (COPD), Squamous cell carcinoma of the cervix, acute myocardial infarction (AMI)/acute heart failure, Chron's Disease, diabetes mellitus type II, Esophageal carcinoma, Squamous cell carcinoma of the larynx, Acute and chronic leukemia of the bone marrow, Lung carcinoma, Malignant lymphoma, Multiple Sclerosis, Ovarian carcinoma, Parkinson disease, Prostate adenocarcinoma, psoriasis, Rheumatoid Arthritis, Renal cell carcinoma, Squamous cell carcinoma of skin, Adenocarcinoma of the stomach, carcinoma of the thyroid gland, Testicular cancer, ulcerative colitis, or Uterine adenocarcinoma.
37 . The method of claim 22 , wherein the disease or disorder comprises a cancer, a premalignant condition, an inflammatory disease, an immune disease, an autoimmune disease or disorder, a cardiovascular disease or disorder, neurological disease or disorder, infectious disease or pain.
38 - 85 . (canceled)
86 . The method of claim 22 , wherein at least one member of the plurality of oligonucleotides comprises at least one functional modification.
87 . A kit comprising at least one reagent for carrying out the method of claim 22 , wherein the at least one reagent comprises the plurality of oligonucleotides.
88 . (canceled)
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90 . (canceled)
91 . The kit of claim 87 , wherein the at least one reagent further comprises at least one of:
a) at least one microvesicle isolation reagent selected from the group consisting of a binding agent to a microvesicle antigen, a column, a substrate, a filtration unit, a polymer, polyethylene glycol, PEG4000, PEG8000, a particle and a bead;
b) at least one oligonucleotide primer for amplifying, sequencing, hybridizing or detecting the plurality of oligonucleotides; and
c) a column or substrate for removing at least one abundant protein from the biological test sample, wherein the at least one abundant protein comprises at least one of albumin, immunoglobulin, fibrinogen and fibrin.
92 - 199 . (canceled)