CN110106229B - 用于分离微囊泡的方法 - Google Patents
用于分离微囊泡的方法 Download PDFInfo
- Publication number
- CN110106229B CN110106229B CN201910000819.0A CN201910000819A CN110106229B CN 110106229 B CN110106229 B CN 110106229B CN 201910000819 A CN201910000819 A CN 201910000819A CN 110106229 B CN110106229 B CN 110106229B
- Authority
- CN
- China
- Prior art keywords
- rna
- microvesicle
- microvesicles
- membrane
- plasma
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 186
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 111
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 111
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 111
- 239000012472 biological sample Substances 0.000 claims abstract description 86
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 334
- 210000002381 plasma Anatomy 0.000 claims description 246
- 239000012528 membrane Substances 0.000 claims description 236
- 108020004999 messenger RNA Proteins 0.000 claims description 114
- 239000012160 loading buffer Substances 0.000 claims description 93
- 239000002679 microRNA Substances 0.000 claims description 76
- 230000007935 neutral effect Effects 0.000 claims description 68
- 210000002700 urine Anatomy 0.000 claims description 24
- 239000011148 porous material Substances 0.000 claims description 20
- 210000001175 cerebrospinal fluid Anatomy 0.000 claims description 19
- 125000001453 quaternary ammonium group Chemical group 0.000 claims description 9
- 210000002966 serum Anatomy 0.000 claims description 9
- 238000004113 cell culture Methods 0.000 claims description 8
- 229920002678 cellulose Polymers 0.000 claims description 8
- 239000012228 culture supernatant Substances 0.000 claims description 8
- 108091070501 miRNA Proteins 0.000 claims description 8
- 230000002934 lysing effect Effects 0.000 claims description 5
- 239000001913 cellulose Substances 0.000 claims description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 320
- 238000005199 ultracentrifugation Methods 0.000 description 164
- 239000011780 sodium chloride Substances 0.000 description 160
- 239000011324 bead Substances 0.000 description 142
- 238000001914 filtration Methods 0.000 description 120
- 239000000523 sample Substances 0.000 description 107
- 238000002955 isolation Methods 0.000 description 97
- 239000000872 buffer Substances 0.000 description 89
- DDSXHBMICAUYKG-UHFFFAOYSA-N 2-[2-[[1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl]amino]propan-2-ylamino]-2-(hydroxymethyl)propane-1,3-diol Chemical compound OCC(NC(C)(C)NC(CO)(CO)CO)(CO)CO DDSXHBMICAUYKG-UHFFFAOYSA-N 0.000 description 88
- 108700011259 MicroRNAs Proteins 0.000 description 88
- 239000011534 wash buffer Substances 0.000 description 88
- 239000004627 regenerated cellulose Substances 0.000 description 76
- 238000000605 extraction Methods 0.000 description 67
- 238000003757 reverse transcription PCR Methods 0.000 description 62
- 108090000623 proteins and genes Proteins 0.000 description 57
- 239000006167 equilibration buffer Substances 0.000 description 56
- 239000000706 filtrate Substances 0.000 description 53
- 238000010804 cDNA synthesis Methods 0.000 description 49
- 108020004635 Complementary DNA Proteins 0.000 description 47
- 239000002299 complementary DNA Substances 0.000 description 47
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 47
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 46
- 239000002245 particle Substances 0.000 description 45
- 238000011067 equilibration Methods 0.000 description 44
- 108020004414 DNA Proteins 0.000 description 43
- 238000002123 RNA extraction Methods 0.000 description 42
- 230000009089 cytolysis Effects 0.000 description 37
- 238000009826 distribution Methods 0.000 description 37
- 101000984753 Homo sapiens Serine/threonine-protein kinase B-raf Proteins 0.000 description 35
- 102100027103 Serine/threonine-protein kinase B-raf Human genes 0.000 description 35
- 108091091807 let-7a stem-loop Proteins 0.000 description 35
- 108091057746 let-7a-4 stem-loop Proteins 0.000 description 35
- 108091028376 let-7a-5 stem-loop Proteins 0.000 description 35
- 108091024393 let-7a-6 stem-loop Proteins 0.000 description 35
- 108091091174 let-7a-7 stem-loop Proteins 0.000 description 35
- 108091032955 Bacterial small RNA Proteins 0.000 description 34
- 230000003321 amplification Effects 0.000 description 34
- 238000004458 analytical method Methods 0.000 description 34
- 238000003199 nucleic acid amplification method Methods 0.000 description 34
- 230000027455 binding Effects 0.000 description 33
- 238000011084 recovery Methods 0.000 description 33
- 239000000203 mixture Substances 0.000 description 32
- 239000007983 Tris buffer Substances 0.000 description 31
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 31
- 230000014509 gene expression Effects 0.000 description 30
- 238000011529 RT qPCR Methods 0.000 description 28
- 108091027943 miR-16 stem-loop Proteins 0.000 description 28
- 239000006228 supernatant Substances 0.000 description 27
- 238000001514 detection method Methods 0.000 description 25
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 23
- 239000003153 chemical reaction reagent Substances 0.000 description 23
- 238000005119 centrifugation Methods 0.000 description 22
- 239000012139 lysis buffer Substances 0.000 description 22
- 238000006243 chemical reaction Methods 0.000 description 21
- 238000011068 loading method Methods 0.000 description 21
- 230000002441 reversible effect Effects 0.000 description 21
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 20
- 206010028980 Neoplasm Diseases 0.000 description 20
- 238000003762 quantitative reverse transcription PCR Methods 0.000 description 20
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 18
- 239000012530 fluid Substances 0.000 description 18
- 230000001965 increasing effect Effects 0.000 description 18
- 238000000746 purification Methods 0.000 description 18
- 238000003828 vacuum filtration Methods 0.000 description 18
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 description 17
- 239000004593 Epoxy Substances 0.000 description 16
- 108091046841 MiR-150 Proteins 0.000 description 16
- 238000002474 experimental method Methods 0.000 description 16
- 239000008188 pellet Substances 0.000 description 16
- 102100029098 Hypoxanthine-guanine phosphoribosyltransferase Human genes 0.000 description 15
- 150000001450 anions Chemical class 0.000 description 15
- 210000001124 body fluid Anatomy 0.000 description 15
- 201000010099 disease Diseases 0.000 description 15
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 15
- 239000003161 ribonuclease inhibitor Substances 0.000 description 15
- 238000000926 separation method Methods 0.000 description 15
- 150000003457 sulfones Chemical class 0.000 description 15
- 229920002554 vinyl polymer Polymers 0.000 description 15
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 14
- 239000000463 material Substances 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 14
- 102100039540 Exocyst complex component 7 Human genes 0.000 description 13
- 101000813489 Homo sapiens Exocyst complex component 7 Proteins 0.000 description 13
- 230000035772 mutation Effects 0.000 description 13
- 238000004448 titration Methods 0.000 description 13
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 12
- 239000012148 binding buffer Substances 0.000 description 12
- 238000010805 cDNA synthesis kit Methods 0.000 description 12
- 230000001143 conditioned effect Effects 0.000 description 12
- 230000000694 effects Effects 0.000 description 12
- 150000003839 salts Chemical class 0.000 description 12
- 101000896557 Homo sapiens Eukaryotic translation initiation factor 3 subunit B Proteins 0.000 description 11
- 101000988834 Homo sapiens Hypoxanthine-guanine phosphoribosyltransferase Proteins 0.000 description 11
- 150000001412 amines Chemical group 0.000 description 11
- 238000003556 assay Methods 0.000 description 11
- 239000000090 biomarker Substances 0.000 description 10
- OWMVSZAMULFTJU-UHFFFAOYSA-N bis-tris Chemical compound OCCN(CCO)C(CO)(CO)CO OWMVSZAMULFTJU-UHFFFAOYSA-N 0.000 description 10
- 238000011534 incubation Methods 0.000 description 10
- 238000000108 ultra-filtration Methods 0.000 description 10
- 201000011510 cancer Diseases 0.000 description 9
- 150000001768 cations Chemical class 0.000 description 9
- 230000005764 inhibitory process Effects 0.000 description 9
- 238000002360 preparation method Methods 0.000 description 9
- 239000000377 silicon dioxide Substances 0.000 description 9
- 238000005406 washing Methods 0.000 description 9
- 241000713887 Human endogenous retrovirus Species 0.000 description 8
- -1 RNA Chemical class 0.000 description 8
- 239000010839 body fluid Substances 0.000 description 8
- 238000003745 diagnosis Methods 0.000 description 8
- 238000010828 elution Methods 0.000 description 8
- 229920000642 polymer Polymers 0.000 description 8
- 230000008569 process Effects 0.000 description 8
- 102200055464 rs113488022 Human genes 0.000 description 8
- 102000035195 Peptidases Human genes 0.000 description 7
- 108091005804 Peptidases Proteins 0.000 description 7
- 239000004365 Protease Substances 0.000 description 7
- 239000000427 antigen Substances 0.000 description 7
- 108091007433 antigens Proteins 0.000 description 7
- 102000036639 antigens Human genes 0.000 description 7
- 239000003599 detergent Substances 0.000 description 7
- 238000010586 diagram Methods 0.000 description 7
- 230000002255 enzymatic effect Effects 0.000 description 7
- 210000001808 exosome Anatomy 0.000 description 7
- 238000003633 gene expression assay Methods 0.000 description 7
- 239000001488 sodium phosphate Substances 0.000 description 7
- 229910000162 sodium phosphate Inorganic materials 0.000 description 7
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 102100027314 Beta-2-microglobulin Human genes 0.000 description 6
- 102100026031 Beta-glucuronidase Human genes 0.000 description 6
- 101000937544 Homo sapiens Beta-2-microglobulin Proteins 0.000 description 6
- 101000933465 Homo sapiens Beta-glucuronidase Proteins 0.000 description 6
- 239000004677 Nylon Substances 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 6
- 239000008280 blood Substances 0.000 description 6
- ZBCBWPMODOFKDW-UHFFFAOYSA-N diethanolamine Chemical compound OCCNCCO ZBCBWPMODOFKDW-UHFFFAOYSA-N 0.000 description 6
- 238000010790 dilution Methods 0.000 description 6
- 239000012895 dilution Substances 0.000 description 6
- 125000000524 functional group Chemical group 0.000 description 6
- 229920001778 nylon Polymers 0.000 description 6
- 238000005191 phase separation Methods 0.000 description 6
- 238000012545 processing Methods 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- 238000003753 real-time PCR Methods 0.000 description 6
- 239000011347 resin Substances 0.000 description 6
- 229920005989 resin Polymers 0.000 description 6
- 238000010839 reverse transcription Methods 0.000 description 6
- 239000013017 sartobind Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 241000894007 species Species 0.000 description 6
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 6
- 102000006382 Ribonucleases Human genes 0.000 description 5
- 108010083644 Ribonucleases Proteins 0.000 description 5
- 230000005856 abnormality Effects 0.000 description 5
- 230000002411 adverse Effects 0.000 description 5
- 239000003729 cation exchange resin Substances 0.000 description 5
- 230000001413 cellular effect Effects 0.000 description 5
- 239000000306 component Substances 0.000 description 5
- 230000001419 dependent effect Effects 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 239000006166 lysate Substances 0.000 description 5
- 238000012544 monitoring process Methods 0.000 description 5
- 238000007781 pre-processing Methods 0.000 description 5
- 238000004393 prognosis Methods 0.000 description 5
- 235000019419 proteases Nutrition 0.000 description 5
- 108020004418 ribosomal RNA Proteins 0.000 description 5
- 241000701161 unidentified adenovirus Species 0.000 description 5
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 4
- 101710132601 Capsid protein Proteins 0.000 description 4
- 101710094648 Coat protein Proteins 0.000 description 4
- 241001534160 Escherichia virus Qbeta Species 0.000 description 4
- 108010091358 Hypoxanthine Phosphoribosyltransferase Proteins 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 101710125418 Major capsid protein Proteins 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 101710141454 Nucleoprotein Proteins 0.000 description 4
- 229910019142 PO4 Inorganic materials 0.000 description 4
- 101710083689 Probable capsid protein Proteins 0.000 description 4
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 description 4
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 230000002378 acidificating effect Effects 0.000 description 4
- 239000003957 anion exchange resin Substances 0.000 description 4
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 4
- 125000002091 cationic group Chemical group 0.000 description 4
- 239000013522 chelant Substances 0.000 description 4
- 230000003750 conditioning effect Effects 0.000 description 4
- 230000002708 enhancing effect Effects 0.000 description 4
- 102000052116 epidermal growth factor receptor activity proteins Human genes 0.000 description 4
- 108700015053 epidermal growth factor receptor activity proteins Proteins 0.000 description 4
- 239000003365 glass fiber Substances 0.000 description 4
- NBZBKCUXIYYUSX-UHFFFAOYSA-N iminodiacetic acid Chemical compound OC(=O)CNCC(O)=O NBZBKCUXIYYUSX-UHFFFAOYSA-N 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 238000005342 ion exchange Methods 0.000 description 4
- 201000001441 melanoma Diseases 0.000 description 4
- 229910052751 metal Inorganic materials 0.000 description 4
- 239000002184 metal Substances 0.000 description 4
- YOHYSYJDKVYCJI-UHFFFAOYSA-N n-[3-[[6-[3-(trifluoromethyl)anilino]pyrimidin-4-yl]amino]phenyl]cyclopropanecarboxamide Chemical compound FC(F)(F)C1=CC=CC(NC=2N=CN=C(NC=3C=C(NC(=O)C4CC4)C=CC=3)C=2)=C1 YOHYSYJDKVYCJI-UHFFFAOYSA-N 0.000 description 4
- 108091027963 non-coding RNA Proteins 0.000 description 4
- 102000042567 non-coding RNA Human genes 0.000 description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 4
- 239000010452 phosphate Substances 0.000 description 4
- 238000011045 prefiltration Methods 0.000 description 4
- 239000012465 retentate Substances 0.000 description 4
- 238000004626 scanning electron microscopy Methods 0.000 description 4
- 238000011282 treatment Methods 0.000 description 4
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 3
- 102100021181 Golgi phosphoprotein 3 Human genes 0.000 description 3
- 101001046870 Homo sapiens Hypoxia-inducible factor 1-alpha Proteins 0.000 description 3
- 102100022875 Hypoxia-inducible factor 1-alpha Human genes 0.000 description 3
- 108020004566 Transfer RNA Proteins 0.000 description 3
- MZVQCMJNVPIDEA-UHFFFAOYSA-N [CH2]CN(CC)CC Chemical group [CH2]CN(CC)CC MZVQCMJNVPIDEA-UHFFFAOYSA-N 0.000 description 3
- 238000001042 affinity chromatography Methods 0.000 description 3
- 125000000129 anionic group Chemical group 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 239000000356 contaminant Substances 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000010195 expression analysis Methods 0.000 description 3
- 230000004907 flux Effects 0.000 description 3
- 238000005227 gel permeation chromatography Methods 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 229920000140 heteropolymer Polymers 0.000 description 3
- 239000003112 inhibitor Substances 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 238000003253 miRNA assay Methods 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 229910000160 potassium phosphate Inorganic materials 0.000 description 3
- 235000011009 potassium phosphates Nutrition 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 230000002485 urinary effect Effects 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- 108091023037 Aptamer Proteins 0.000 description 2
- 206010003445 Ascites Diseases 0.000 description 2
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 102000016911 Deoxyribonucleases Human genes 0.000 description 2
- 108010053770 Deoxyribonucleases Proteins 0.000 description 2
- 229920002307 Dextran Polymers 0.000 description 2
- 102000018651 Epithelial Cell Adhesion Molecule Human genes 0.000 description 2
- 108010066687 Epithelial Cell Adhesion Molecule Proteins 0.000 description 2
- 241000282326 Felis catus Species 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 102000005431 Molecular Chaperones Human genes 0.000 description 2
- 108010006519 Molecular Chaperones Proteins 0.000 description 2
- 229920001213 Polysorbate 20 Polymers 0.000 description 2
- 206010036790 Productive cough Diseases 0.000 description 2
- 238000010802 RNA extraction kit Methods 0.000 description 2
- 239000013614 RNA sample Substances 0.000 description 2
- 238000010240 RT-PCR analysis Methods 0.000 description 2
- 108020004511 Recombinant DNA Proteins 0.000 description 2
- 241000283984 Rodentia Species 0.000 description 2
- 108020004459 Small interfering RNA Proteins 0.000 description 2
- 239000013504 Triton X-100 Substances 0.000 description 2
- 229920004890 Triton X-100 Polymers 0.000 description 2
- 210000004381 amniotic fluid Anatomy 0.000 description 2
- 238000005349 anion exchange Methods 0.000 description 2
- 210000000601 blood cell Anatomy 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 229940023913 cation exchange resins Drugs 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 210000002726 cyst fluid Anatomy 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000001085 differential centrifugation Methods 0.000 description 2
- 238000002270 exclusion chromatography Methods 0.000 description 2
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 description 2
- 238000005194 fractionation Methods 0.000 description 2
- 235000020256 human milk Nutrition 0.000 description 2
- 210000004251 human milk Anatomy 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000003014 ion exchange membrane Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000002751 lymph Anatomy 0.000 description 2
- 210000004324 lymphatic system Anatomy 0.000 description 2
- 238000002826 magnetic-activated cell sorting Methods 0.000 description 2
- 230000003211 malignant effect Effects 0.000 description 2
- 125000000250 methylamino group Chemical group [H]N(*)C([H])([H])[H] 0.000 description 2
- ZIYVHBGGAOATLY-UHFFFAOYSA-N methylmalonic acid Chemical compound OC(=O)C(C)C(O)=O ZIYVHBGGAOATLY-UHFFFAOYSA-N 0.000 description 2
- 239000012982 microporous membrane Substances 0.000 description 2
- 231100000150 mutagenicity / genotoxicity testing Toxicity 0.000 description 2
- 210000002445 nipple Anatomy 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 239000013610 patient sample Substances 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 210000004910 pleural fluid Anatomy 0.000 description 2
- 229920002492 poly(sulfone) Polymers 0.000 description 2
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 2
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 2
- 239000001294 propane Substances 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 210000000582 semen Anatomy 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000004055 small Interfering RNA Substances 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 210000003802 sputum Anatomy 0.000 description 2
- 208000024794 sputum Diseases 0.000 description 2
- 239000013595 supernatant sample Substances 0.000 description 2
- 210000001138 tear Anatomy 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- NLMKTBGFQGKQEV-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-hexadecoxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CCCCCCCCCCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO NLMKTBGFQGKQEV-UHFFFAOYSA-N 0.000 description 1
- IEQAICDLOKRSRL-UHFFFAOYSA-N 2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-[2-(2-dodecoxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethanol Chemical compound CCCCCCCCCCCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO IEQAICDLOKRSRL-UHFFFAOYSA-N 0.000 description 1
- UMCMPZBLKLEWAF-BCTGSCMUSA-N 3-[(3-cholamidopropyl)dimethylammonio]propane-1-sulfonate Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCC[N+](C)(C)CCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 UMCMPZBLKLEWAF-BCTGSCMUSA-N 0.000 description 1
- GUQQBLRVXOUDTN-XOHPMCGNSA-N 3-[dimethyl-[3-[[(4r)-4-[(3r,5s,7r,8r,9s,10s,12s,13r,14s,17r)-3,7,12-trihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]pentanoyl]amino]propyl]azaniumyl]-2-hydroxypropane-1-sulfonate Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCC[N+](C)(C)CC(O)CS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 GUQQBLRVXOUDTN-XOHPMCGNSA-N 0.000 description 1
- 101710159080 Aconitate hydratase A Proteins 0.000 description 1
- 101710159078 Aconitate hydratase B Proteins 0.000 description 1
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 241000700199 Cavia porcellus Species 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 238000007399 DNA isolation Methods 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 241000283073 Equus caballus Species 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 101000914484 Homo sapiens T-lymphocyte activation antigen CD80 Proteins 0.000 description 1
- 108091069019 Homo sapiens miR-124-1 stem-loop Proteins 0.000 description 1
- 108091069008 Homo sapiens miR-124-2 stem-loop Proteins 0.000 description 1
- 108091069007 Homo sapiens miR-124-3 stem-loop Proteins 0.000 description 1
- 108091068993 Homo sapiens miR-142 stem-loop Proteins 0.000 description 1
- 238000009015 Human TaqMan MicroRNA Assay kit Methods 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 101150105104 Kras gene Proteins 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- 108020005196 Mitochondrial DNA Proteins 0.000 description 1
- 108010085220 Multiprotein Complexes Proteins 0.000 description 1
- 102000007474 Multiprotein Complexes Human genes 0.000 description 1
- 206010051141 Myeloblastoma Diseases 0.000 description 1
- 108700020796 Oncogene Proteins 0.000 description 1
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 1
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 description 1
- 230000007022 RNA scission Effects 0.000 description 1
- 102000044126 RNA-Binding Proteins Human genes 0.000 description 1
- 230000004570 RNA-binding Effects 0.000 description 1
- 101710105008 RNA-binding protein Proteins 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 102000004389 Ribonucleoproteins Human genes 0.000 description 1
- 108010081734 Ribonucleoproteins Proteins 0.000 description 1
- 239000012506 Sephacryl® Substances 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 239000012505 Superdex™ Substances 0.000 description 1
- 102100027222 T-lymphocyte activation antigen CD80 Human genes 0.000 description 1
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 1
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 1
- 108010067390 Viral Proteins Proteins 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 125000003172 aldehyde group Chemical group 0.000 description 1
- 238000007844 allele-specific PCR Methods 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000001640 apoptogenic effect Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 239000013060 biological fluid Substances 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 239000012503 blood component Substances 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 235000011089 carbon dioxide Nutrition 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 235000013330 chicken meat Nutrition 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000008711 chromosomal rearrangement Effects 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 229920001577 copolymer Polymers 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 1
- 125000001664 diethylamino group Chemical group [H]C([H])([H])C([H])([H])N(*)C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000012149 elution buffer Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000012869 ethanol precipitation Methods 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000028023 exocytosis Effects 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 239000010200 folin Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 208000005017 glioblastoma Diseases 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000003394 haemopoietic effect Effects 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 239000012510 hollow fiber Substances 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 235000013847 iso-butane Nutrition 0.000 description 1
- NNPPMTNAJDCUHE-UHFFFAOYSA-N isobutane Chemical compound CC(C)C NNPPMTNAJDCUHE-UHFFFAOYSA-N 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000006148 magnetic separator Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 108091034121 miR-92a stem-loop Proteins 0.000 description 1
- 108091041519 miR-92a-3 stem-loop Proteins 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 238000001823 molecular biology technique Methods 0.000 description 1
- 229920000344 molecularly imprinted polymer Polymers 0.000 description 1
- 210000002487 multivesicular body Anatomy 0.000 description 1
- CGVLVOOFCGWBCS-RGDJUOJXSA-N n-octyl β-d-thioglucopyranoside Chemical compound CCCCCCCCS[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O CGVLVOOFCGWBCS-RGDJUOJXSA-N 0.000 description 1
- 210000003739 neck Anatomy 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 238000007826 nucleic acid assay Methods 0.000 description 1
- HEGSGKPQLMEBJL-RKQHYHRCSA-N octyl beta-D-glucopyranoside Chemical compound CCCCCCCCO[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HEGSGKPQLMEBJL-RKQHYHRCSA-N 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 210000004789 organ system Anatomy 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 238000000710 polymer precipitation Methods 0.000 description 1
- 102000054765 polymorphisms of proteins Human genes 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 210000002307 prostate Anatomy 0.000 description 1
- 230000009145 protein modification Effects 0.000 description 1
- 239000013014 purified material Substances 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 210000003705 ribosome Anatomy 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- IJDNQMDRQITEOD-UHFFFAOYSA-N sec-butylidene Natural products CCCC IJDNQMDRQITEOD-UHFFFAOYSA-N 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 238000012204 single target assay Methods 0.000 description 1
- 238000001542 size-exclusion chromatography Methods 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000012799 strong cation exchange Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 150000003871 sulfonates Chemical group 0.000 description 1
- 125000000542 sulfonic acid group Chemical group 0.000 description 1
- 239000012134 supernatant fraction Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 230000009452 underexpressoin Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
- C12N15/1017—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by filtration, e.g. using filters, frits, membranes
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Crystallography & Structural Chemistry (AREA)
- Plant Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201361748575P | 2013-01-03 | 2013-01-03 | |
| US61/748575 | 2013-01-03 | ||
| PCT/US2014/010173 WO2014107571A1 (en) | 2013-01-03 | 2014-01-03 | Methods for isolating microvesicles |
| CN201480002901.2A CN105026911B (zh) | 2013-01-03 | 2014-01-03 | 用于分离微囊泡的方法 |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201480002901.2A Division CN105026911B (zh) | 2013-01-03 | 2014-01-03 | 用于分离微囊泡的方法 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110106229A CN110106229A (zh) | 2019-08-09 |
| CN110106229B true CN110106229B (zh) | 2023-03-28 |
Family
ID=51062472
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910000819.0A Active CN110106229B (zh) | 2013-01-03 | 2014-01-03 | 用于分离微囊泡的方法 |
| CN201480002901.2A Active CN105026911B (zh) | 2013-01-03 | 2014-01-03 | 用于分离微囊泡的方法 |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201480002901.2A Active CN105026911B (zh) | 2013-01-03 | 2014-01-03 | 用于分离微囊泡的方法 |
Country Status (10)
| Country | Link |
|---|---|
| US (2) | US20150353920A1 (enExample) |
| EP (2) | EP3499212B1 (enExample) |
| JP (1) | JP6449781B2 (enExample) |
| CN (2) | CN110106229B (enExample) |
| AU (2) | AU2014203987B2 (enExample) |
| BR (1) | BR112015016136A2 (enExample) |
| CA (1) | CA2897207C (enExample) |
| IL (2) | IL239786B (enExample) |
| MX (2) | MX366153B (enExample) |
| WO (1) | WO2014107571A1 (enExample) |
Families Citing this family (57)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3569607A1 (en) * | 2011-05-11 | 2019-11-20 | Exosome Diagnostics, Inc. | Nucleic acid extraction from heterogeneous biological materials |
| ES2981185T3 (es) | 2013-03-14 | 2024-10-07 | Translate Bio Inc | Métodos para la purificación de ARN mensajero |
| US20170165334A1 (en) * | 2015-12-11 | 2017-06-15 | Tianxin Wang | Methods to Treat Diseases with Protein, Peptide, Antigen Modification and Hemopurification |
| US11268085B2 (en) | 2014-05-27 | 2022-03-08 | Exosome Diagnostics, Inc. | Methods for isolating microvesicles and extracting nucleic acids from biological samples |
| CA2954576C (en) | 2014-07-09 | 2020-08-11 | Daniel ENDERLE | Methods for isolating microvesicles and extracting nucleic acids from biological samples |
| US20170211139A1 (en) * | 2014-07-11 | 2017-07-27 | Exosome Diagnostics, Inc. | Compositions and methods for detecting rare sequence variants in nucleic acid sequencing |
| US9783799B2 (en) * | 2014-10-24 | 2017-10-10 | Abbott Molecular Inc. | Enrichment of small nucleic acids |
| EP3112474A1 (en) | 2015-06-29 | 2017-01-04 | Evonik Degussa GmbH | Method of detecting avian necrotic enteritis |
| WO2017062901A2 (en) * | 2015-10-07 | 2017-04-13 | Tymora Analytical Operations Llc | Methods to isolate extracellular vesicles in biofluids for biomarker discovery and disease diagnosis |
| KR20190020649A (ko) | 2016-04-15 | 2019-03-04 | 엑소좀 디아그노스틱스, 인크. | 역형성 림프종 키나아제(alk) 핵산 및 alk 융합 전사물의 혈장 기반 검출 및 암의 진단 및 치료에 있어 이의 용도 |
| CN105716923A (zh) * | 2016-04-21 | 2016-06-29 | 华南理工大学 | 一种聚合物囊泡扫描电镜样品的制备方法 |
| US11214833B2 (en) | 2016-05-05 | 2022-01-04 | Exosome Diagnostics, Inc. | Profiling microvesicle nucleic acids and uses thereof as signatures in diagnosis of renal transplant rejection |
| WO2017197399A1 (en) * | 2016-05-13 | 2017-11-16 | Exosome Diagnostics, Inc. | Automated and manual methods for isolation of extracellular vesicles and co-isolation of cell-free dna from biofluids |
| US11614388B2 (en) | 2016-10-13 | 2023-03-28 | H.U. Group Research Institute G.K. | Method for recovering extracellular vesicles |
| CN110191962A (zh) * | 2016-10-21 | 2019-08-30 | 外来体诊断公司 | 外来体相关核酸的测序和分析 |
| AU2017366813B2 (en) | 2016-11-30 | 2023-04-20 | Exosome Diagnostics, Inc. | Methods and compositions to detect mutations in plasma using exosomal RNA and cell free DNA from non-small cell lung cancer patients |
| WO2018112557A1 (en) * | 2016-12-23 | 2018-06-28 | Altnia Operations Pty Ltd | Methods and compositions for purification or isolation of microvesicles and exosomes |
| WO2018126278A2 (en) | 2017-01-02 | 2018-07-05 | Exosome Diagnostics, Inc. | Methods to distinguish rna and dna in a combined preparation |
| KR101875594B1 (ko) * | 2017-01-13 | 2018-07-06 | ㈜로제타엑소좀 | 금속을 이용한 세포밖 소포체의 분리 방법 |
| DK3622082T3 (da) | 2017-05-12 | 2023-06-12 | Evonik Operations Gmbh | Fremgangsmåde til påvisning af c. perfringens-inducerede sygdomme i en fugleflok |
| CN110945145B (zh) | 2017-05-17 | 2024-06-11 | 外来体诊断公司 | 微泡核酸和/或蛋白及其作为肾移植排斥的标志物的应用 |
| CN107271655A (zh) * | 2017-05-18 | 2017-10-20 | 成都中医药大学附属医院 | 一种检测尿外泌体荷载miRNAs的试剂盒和方法 |
| US11390864B2 (en) * | 2017-07-12 | 2022-07-19 | Illumina, Inc. | Nucleic acid extraction materials, systems, and methods |
| EP3652315A4 (en) | 2017-07-12 | 2021-09-01 | Exosome Diagnostics, Inc. | METHODS FOR ISOLATION AND ENRICHMENT OF EXTRACELLULAR VESICLE POPULATIONS DERIVED FROM BIOFLUIDS, AND ASSOCIATED METHODS OF USE |
| EP3655531B1 (en) * | 2017-07-18 | 2024-09-04 | Exosome Diagnostics, Inc. | Sequencing of nucleic acids associated with exosomal isolation from patients with glioblastoma multiforme |
| US11904259B2 (en) * | 2017-07-26 | 2024-02-20 | Rosetta Exosome | Method for isolating extracellular vesicles using cations |
| RU2745613C1 (ru) * | 2017-12-26 | 2021-03-29 | Общество с ограниченной ответственностью "Простагност" (ООО "Простагност") | Способ и устройство для выделения внеклеточных везикул из биологических жидкостей с помощью каскадной ультрафильтрации |
| CA3092472A1 (en) | 2018-03-02 | 2019-09-06 | Evonik Operations Gmbh | In vitro method for detecting intestinal barrier failure in animals |
| US12378608B2 (en) | 2018-06-06 | 2025-08-05 | Exosome Diagnostics, Inc. | Methods for developing urine biomarkers and for detecting bladder cancer |
| US12161950B2 (en) | 2018-06-18 | 2024-12-10 | Exopharm Limited | Methods and compositions for purification or isolation of microvesicles and exosomes |
| CN108841777A (zh) * | 2018-06-22 | 2018-11-20 | 北京恩泽康泰生物科技有限公司 | 基于静电吸附的胞外囊泡及其内含物的提取方法及装置 |
| US20210239685A1 (en) * | 2018-06-27 | 2021-08-05 | The Trustees Of Indiana University | Methods of analyzing dna in urine |
| US10955379B2 (en) | 2018-09-27 | 2021-03-23 | Taiwan Semiconductor Manufacturing Co., Ltd. | Differential sensing with BioFET sensors |
| CN109337859A (zh) * | 2018-10-08 | 2019-02-15 | 南昌大学 | 一种分离细胞表面囊泡的方法 |
| JP7426725B2 (ja) * | 2018-10-30 | 2024-02-02 | Craif株式会社 | 細胞外小胞を捕捉するために用いられるデバイス、細胞外小胞の保存方法および移送方法 |
| WO2020106853A1 (en) | 2018-11-20 | 2020-05-28 | Exosome Diagnostics, Inc. | Compositions and methods for internal controls of microvesicle isolations |
| JP2020092688A (ja) * | 2018-12-12 | 2020-06-18 | 国立大学法人東海国立大学機構 | マイクロrnaを含む体液抽出物 |
| ES2938227T3 (es) | 2018-12-14 | 2023-04-05 | Evonik Operations Gmbh | Método in vitro de detección de disbiosis intestinal aviar |
| EP3998349A3 (en) | 2019-03-01 | 2022-08-24 | Mercy Bioanalytics, Inc. | Compositions and methods for target protein detection on exosomes using proximal ligation |
| US12233356B2 (en) | 2019-03-21 | 2025-02-25 | Lonza Sales Ag | Process for preparing extracellular vesicles |
| CN112048462B (zh) * | 2019-06-05 | 2022-08-23 | 北京丰特云基科技发展有限公司 | 一种基于阴离子多聚物修饰基质的细胞外囊泡分离富集方法 |
| WO2021055770A2 (en) | 2019-09-18 | 2021-03-25 | Exosome Diagnostics, Inc. | Compositions, methods, and kits for the isolation of extracellular vesicles |
| US10984211B1 (en) | 2019-10-18 | 2021-04-20 | Taiwan Semiconductor Manufacturing Co., Ltd. | Semiconductor device with bioFET and biometric sensors |
| CN114787349A (zh) | 2019-12-16 | 2022-07-22 | 凯杰有限公司 | 富集方法 |
| US20230009972A1 (en) | 2019-12-16 | 2023-01-12 | Qiagen Gmbh | Method for enriching vesicular rna |
| JP7425403B2 (ja) * | 2020-02-17 | 2024-01-31 | 株式会社Jvcケンウッド | 生体試料分析方法 |
| CN113495029A (zh) * | 2020-04-01 | 2021-10-12 | 杨昆德 | 用于分离欲求物质的装置和方法 |
| US20230168162A1 (en) * | 2020-04-24 | 2023-06-01 | Korea University Research And Business Foundation | Microvesicle isolation method and microvesicle isolation |
| US20230203587A1 (en) | 2020-05-29 | 2023-06-29 | Exosome Diagnostics, Inc. | Use of microvesicle signature for the diagnosis and treatment of kidney transplant rejection |
| WO2022066934A2 (en) * | 2020-09-23 | 2022-03-31 | Codiak Biosciences, Inc. | Process for preparing extracellular vesicles |
| CN112391382B (zh) * | 2020-12-07 | 2022-10-11 | 湖北盛齐安生物科技股份有限公司 | 一种快速提取囊泡dna的方法 |
| JP7773765B2 (ja) * | 2020-12-18 | 2025-11-20 | 国立研究開発法人産業技術総合研究所 | 回収用キットおよび核酸含有溶液の調製方法 |
| US20250075258A1 (en) | 2021-07-16 | 2025-03-06 | Exosome Diagnostics, Inc. | Methods of detecting sjogren's syndrome using salivary exosomes |
| KR20240161644A (ko) | 2022-02-18 | 2024-11-12 | 엑소좀 디아그노스틱스, 인크. | 신장 장애의 식별 및 치료에 있어서의 미세 소포 시그니처의 용도 |
| WO2024054572A1 (en) | 2022-09-07 | 2024-03-14 | Exosome Diagnostics, Inc. | Methods of detecting sjögren's syndrome using salivary exosomes |
| WO2025057787A1 (ja) * | 2023-09-14 | 2025-03-20 | 東洋紡株式会社 | 細胞外小胞の製造方法 |
| CN117487739B (zh) * | 2023-10-30 | 2024-10-01 | 深圳市茵冠生物科技有限公司 | 一种大规模纯化293细胞外泌体的方法 |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5438128A (en) * | 1992-02-07 | 1995-08-01 | Millipore Corporation | Method for rapid purifiction of nucleic acids using layered ion-exchange membranes |
| JP2004105009A (ja) * | 2002-09-13 | 2004-04-08 | Dojindo Laboratories | テトラフェニルホウ素化合物から成る核酸分離用試薬とそれを用いる核酸分離方法 |
| WO2012087241A1 (en) * | 2010-12-20 | 2012-06-28 | Agency For Science, Technology And Research | Method of purifying exosomes |
| CN102596177A (zh) * | 2009-07-01 | 2012-07-18 | 阿昂梅迪克斯公司 | 来源于有核哺乳动物细胞的微囊泡及其应用 |
| WO2012174282A2 (en) * | 2011-06-16 | 2012-12-20 | Caris Life Sciences Luxembourg Holdings, S.A.R.L. | Biomarker compositions and methods |
Family Cites Families (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4935342A (en) * | 1986-12-01 | 1990-06-19 | Syngene, Inc. | Method of isolating and purifying nucleic acids from biological samples |
| US5639606A (en) | 1993-04-06 | 1997-06-17 | The University Of Rochester | Method for quantitative measurement of gene expression using multiplex competitive reverse transcriptase-polymerase chain reaction |
| FR2788780B1 (fr) | 1999-01-27 | 2001-03-30 | Ap Cells Inc | Procede de preparation de vesicules membranaires |
| GB9927320D0 (en) | 1999-11-18 | 2000-01-12 | Chiron Spa | Exosome separation |
| US6812023B1 (en) | 2000-04-27 | 2004-11-02 | Anosys, Inc. | Methods of producing membrane vesicles |
| EP2604704B1 (en) | 2008-02-01 | 2018-10-03 | The General Hospital Corporation | Use of microvesicles in diagnosis and prognosis of brain tumor |
| WO2010065765A2 (en) * | 2008-12-04 | 2010-06-10 | Aethlon Medical, Inc. | Affinity capture of circulating biomarkers |
| US20120077263A1 (en) * | 2009-06-05 | 2012-03-29 | Mayo Foundation For Medical Education And Research | Methods and materials for isolating exosomes |
| WO2012006476A2 (en) * | 2010-07-07 | 2012-01-12 | Aethlon Medical, Inc. | Methods and compositions for quantifying exosomes |
| EP2638057B1 (en) * | 2010-11-10 | 2019-03-06 | Exosome Diagnostics, Inc. | Method for isolation of nucleic acid containing particles and extraction of nucleic acids therefrom |
-
2014
- 2014-01-03 BR BR112015016136A patent/BR112015016136A2/pt not_active IP Right Cessation
- 2014-01-03 JP JP2015551774A patent/JP6449781B2/ja active Active
- 2014-01-03 EP EP18199291.8A patent/EP3499212B1/en active Active
- 2014-01-03 EP EP14735397.3A patent/EP2941629B1/en active Active
- 2014-01-03 AU AU2014203987A patent/AU2014203987B2/en active Active
- 2014-01-03 CN CN201910000819.0A patent/CN110106229B/zh active Active
- 2014-01-03 CA CA2897207A patent/CA2897207C/en active Active
- 2014-01-03 WO PCT/US2014/010173 patent/WO2014107571A1/en not_active Ceased
- 2014-01-03 MX MX2015008727A patent/MX366153B/es active IP Right Grant
- 2014-01-03 CN CN201480002901.2A patent/CN105026911B/zh active Active
- 2014-01-03 US US14/759,238 patent/US20150353920A1/en not_active Abandoned
- 2014-01-03 MX MX2019007742A patent/MX392552B/es unknown
-
2015
- 2015-07-05 IL IL239786A patent/IL239786B/en active IP Right Grant
-
2018
- 2018-04-20 AU AU2018202789A patent/AU2018202789B2/en active Active
-
2019
- 2019-06-19 IL IL267526A patent/IL267526B/en active IP Right Grant
-
2020
- 2020-06-05 US US16/893,966 patent/US20210171934A1/en not_active Abandoned
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5438128A (en) * | 1992-02-07 | 1995-08-01 | Millipore Corporation | Method for rapid purifiction of nucleic acids using layered ion-exchange membranes |
| JP2004105009A (ja) * | 2002-09-13 | 2004-04-08 | Dojindo Laboratories | テトラフェニルホウ素化合物から成る核酸分離用試薬とそれを用いる核酸分離方法 |
| CN102596177A (zh) * | 2009-07-01 | 2012-07-18 | 阿昂梅迪克斯公司 | 来源于有核哺乳动物细胞的微囊泡及其应用 |
| WO2012087241A1 (en) * | 2010-12-20 | 2012-06-28 | Agency For Science, Technology And Research | Method of purifying exosomes |
| WO2012174282A2 (en) * | 2011-06-16 | 2012-12-20 | Caris Life Sciences Luxembourg Holdings, S.A.R.L. | Biomarker compositions and methods |
Also Published As
| Publication number | Publication date |
|---|---|
| CA2897207C (en) | 2022-08-02 |
| IL239786B (en) | 2019-07-31 |
| AU2018202789A1 (en) | 2018-05-10 |
| EP3499212B1 (en) | 2020-08-12 |
| US20210171934A1 (en) | 2021-06-10 |
| BR112015016136A2 (pt) | 2017-07-11 |
| IL267526A (en) | 2019-08-29 |
| JP2016502862A (ja) | 2016-02-01 |
| WO2014107571A1 (en) | 2014-07-10 |
| AU2014203987B2 (en) | 2018-01-25 |
| CN105026911B (zh) | 2019-01-22 |
| EP2941629B1 (en) | 2018-10-10 |
| MX2015008727A (es) | 2016-04-21 |
| CN110106229A (zh) | 2019-08-09 |
| AU2014203987A1 (en) | 2015-08-20 |
| MX366153B (es) | 2019-06-28 |
| AU2018202789B2 (en) | 2020-01-02 |
| EP3499212A1 (en) | 2019-06-19 |
| EP2941629A4 (en) | 2017-03-22 |
| MX2019007742A (es) | 2019-08-29 |
| JP6449781B2 (ja) | 2019-01-09 |
| CA2897207A1 (en) | 2014-07-10 |
| IL239786A0 (en) | 2015-08-31 |
| HK1217365A1 (en) | 2017-01-06 |
| EP2941629A1 (en) | 2015-11-11 |
| IL267526B (en) | 2020-10-29 |
| MX392552B (es) | 2025-03-11 |
| CN105026911A (zh) | 2015-11-04 |
| US20150353920A1 (en) | 2015-12-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110106229B (zh) | 用于分离微囊泡的方法 | |
| AU2019250221B2 (en) | Methods for isolating microvesicles and extracting nucleic acids from biological samples | |
| US20210238582A1 (en) | Automated and manual methods for isolation of extracellular vesicles and co-isolation of cell-free dna from biofluids | |
| US11268085B2 (en) | Methods for isolating microvesicles and extracting nucleic acids from biological samples | |
| HK40009837A (en) | Methods for isolating microvesicles | |
| HK1217365B (en) | Methods for isolating microvesicles |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |