DK2970948T3 - Fremgangsmåder til rna-oprensning - Google Patents
Fremgangsmåder til rna-oprensning Download PDFInfo
- Publication number
- DK2970948T3 DK2970948T3 DK14709704.2T DK14709704T DK2970948T3 DK 2970948 T3 DK2970948 T3 DK 2970948T3 DK 14709704 T DK14709704 T DK 14709704T DK 2970948 T3 DK2970948 T3 DK 2970948T3
- Authority
- DK
- Denmark
- Prior art keywords
- rna
- sample
- buffer
- purification
- chromatography
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims description 137
- 238000000746 purification Methods 0.000 title description 114
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims description 387
- 239000000872 buffer Substances 0.000 claims description 100
- 108090000623 proteins and genes Proteins 0.000 claims description 82
- 102000004169 proteins and genes Human genes 0.000 claims description 81
- 238000009295 crossflow filtration Methods 0.000 claims description 80
- 239000011324 bead Substances 0.000 claims description 57
- 239000002773 nucleotide Substances 0.000 claims description 55
- 125000003729 nucleotide group Chemical group 0.000 claims description 55
- 238000013518 transcription Methods 0.000 claims description 51
- 230000035897 transcription Effects 0.000 claims description 51
- 238000006243 chemical reaction Methods 0.000 claims description 50
- 239000012528 membrane Substances 0.000 claims description 50
- 238000000338 in vitro Methods 0.000 claims description 48
- 238000011062 flow through chromatography Methods 0.000 claims description 38
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical group [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 30
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 28
- 230000008569 process Effects 0.000 claims description 24
- 239000008194 pharmaceutical composition Substances 0.000 claims description 19
- 229910000160 potassium phosphate Inorganic materials 0.000 claims description 15
- 235000011009 potassium phosphates Nutrition 0.000 claims description 15
- 239000001103 potassium chloride Substances 0.000 claims description 14
- 235000011164 potassium chloride Nutrition 0.000 claims description 14
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 claims description 12
- 238000004519 manufacturing process Methods 0.000 claims description 11
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 claims description 10
- 108020004999 messenger RNA Proteins 0.000 claims description 10
- 239000001226 triphosphate Substances 0.000 claims description 10
- 235000011178 triphosphate Nutrition 0.000 claims description 10
- 230000005526 G1 to G0 transition Effects 0.000 claims description 9
- 239000000463 material Substances 0.000 claims description 9
- 239000003960 organic solvent Substances 0.000 claims description 9
- 239000002342 ribonucleoside Substances 0.000 claims description 8
- -1 ribonucleoside triphosphates Chemical class 0.000 claims description 7
- 150000004712 monophosphates Chemical class 0.000 claims description 5
- 239000013612 plasmid Substances 0.000 claims description 5
- 239000012634 fragment Substances 0.000 claims description 4
- 210000000170 cell membrane Anatomy 0.000 claims 1
- 230000007515 enzymatic degradation Effects 0.000 claims 1
- 239000000523 sample Substances 0.000 description 118
- 235000018102 proteins Nutrition 0.000 description 75
- 238000004587 chromatography analysis Methods 0.000 description 46
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 43
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 43
- 150000003839 salts Chemical class 0.000 description 42
- 239000000203 mixture Substances 0.000 description 39
- 238000010828 elution Methods 0.000 description 27
- 239000011148 porous material Substances 0.000 description 24
- 239000002245 particle Substances 0.000 description 23
- 238000011084 recovery Methods 0.000 description 23
- 239000003446 ligand Substances 0.000 description 21
- 102000004190 Enzymes Human genes 0.000 description 20
- 108090000790 Enzymes Proteins 0.000 description 20
- 239000012535 impurity Substances 0.000 description 20
- 239000000356 contaminant Substances 0.000 description 19
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 16
- 241000894007 species Species 0.000 description 16
- 239000007983 Tris buffer Substances 0.000 description 15
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 15
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 14
- 230000000694 effects Effects 0.000 description 13
- 230000002163 immunogen Effects 0.000 description 13
- 239000011159 matrix material Substances 0.000 description 12
- 229960005486 vaccine Drugs 0.000 description 12
- 239000012149 elution buffer Substances 0.000 description 10
- 230000000717 retained effect Effects 0.000 description 10
- 239000012465 retentate Substances 0.000 description 10
- 241000710929 Alphavirus Species 0.000 description 9
- 238000009472 formulation Methods 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 229910019142 PO4 Inorganic materials 0.000 description 8
- 239000010452 phosphate Substances 0.000 description 8
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 8
- 239000011780 sodium chloride Substances 0.000 description 8
- 102000016911 Deoxyribonucleases Human genes 0.000 description 7
- 108010053770 Deoxyribonucleases Proteins 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 239000012895 dilution Substances 0.000 description 7
- 238000010790 dilution Methods 0.000 description 7
- 239000000706 filtrate Substances 0.000 description 7
- 108010026228 mRNA guanylyltransferase Proteins 0.000 description 7
- 238000001556 precipitation Methods 0.000 description 7
- 238000011002 quantification Methods 0.000 description 7
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 6
- 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 6
- 238000002965 ELISA Methods 0.000 description 6
- RTYZCUMXOXNVSI-UHFFFAOYSA-N OOOOOOOOOOOOOOOOOO Chemical compound OOOOOOOOOOOOOOOOOO RTYZCUMXOXNVSI-UHFFFAOYSA-N 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000011109 contamination Methods 0.000 description 6
- 239000012530 fluid Substances 0.000 description 6
- 239000008363 phosphate buffer Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 208000002109 Argyria Diseases 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 5
- 239000004695 Polyether sulfone Substances 0.000 description 5
- 239000013614 RNA sample Substances 0.000 description 5
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- 230000008901 benefit Effects 0.000 description 5
- 239000003085 diluting agent Substances 0.000 description 5
- 238000002296 dynamic light scattering Methods 0.000 description 5
- 239000000835 fiber Substances 0.000 description 5
- 238000001914 filtration Methods 0.000 description 5
- 229920006393 polyether sulfone Polymers 0.000 description 5
- 230000002829 reductive effect Effects 0.000 description 5
- 239000012898 sample dilution Substances 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- 239000001488 sodium phosphate Substances 0.000 description 5
- 229910000162 sodium phosphate Inorganic materials 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000001228 spectrum Methods 0.000 description 5
- UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 description 5
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 5
- 108700026244 Open Reading Frames Proteins 0.000 description 4
- 102000006382 Ribonucleases Human genes 0.000 description 4
- 108010083644 Ribonucleases Proteins 0.000 description 4
- 108091027544 Subgenomic mRNA Proteins 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 238000011210 chromatographic step Methods 0.000 description 4
- 238000013461 design Methods 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 235000011180 diphosphates Nutrition 0.000 description 4
- 239000003480 eluent Substances 0.000 description 4
- 239000012467 final product Substances 0.000 description 4
- 230000028993 immune response Effects 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 230000003993 interaction Effects 0.000 description 4
- 230000000069 prophylactic effect Effects 0.000 description 4
- 239000003161 ribonuclease inhibitor Substances 0.000 description 4
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 3
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 3
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 108010009413 Pyrophosphatases Proteins 0.000 description 3
- 102000009609 Pyrophosphatases Human genes 0.000 description 3
- 108020005093 RNA Precursors Proteins 0.000 description 3
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000002738 chelating agent Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 239000007857 degradation product Substances 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000013020 final formulation Substances 0.000 description 3
- 230000004907 flux Effects 0.000 description 3
- 239000012537 formulation buffer Substances 0.000 description 3
- 230000002209 hydrophobic effect Effects 0.000 description 3
- 238000002649 immunization Methods 0.000 description 3
- 238000011065 in-situ storage Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000007918 intramuscular administration Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 229920002492 poly(sulfone) Polymers 0.000 description 3
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 3
- 238000004007 reversed phase HPLC Methods 0.000 description 3
- 229910052709 silver Inorganic materials 0.000 description 3
- 239000004332 silver Substances 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 239000007921 spray Substances 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 238000013519 translation Methods 0.000 description 3
- OGHAROSJZRTIOK-KQYNXXCUSA-O 7-methylguanosine Chemical compound C1=2N=C(N)NC(=O)C=2[N+](C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OGHAROSJZRTIOK-KQYNXXCUSA-O 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical group [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 2
- 101710094648 Coat protein Proteins 0.000 description 2
- 241000124008 Mammalia Species 0.000 description 2
- 108060004795 Methyltransferase Proteins 0.000 description 2
- 108091028664 Ribonucleotide Proteins 0.000 description 2
- 101710172711 Structural protein Proteins 0.000 description 2
- 241000710959 Venezuelan equine encephalitis virus Species 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 150000003973 alkyl amines Chemical class 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 230000027455 binding Effects 0.000 description 2
- 230000008827 biological function Effects 0.000 description 2
- 239000000337 buffer salt Substances 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 2
- 235000019846 buffering salt Nutrition 0.000 description 2
- 150000001768 cations Chemical class 0.000 description 2
- 239000012501 chromatography medium Substances 0.000 description 2
- 239000007979 citrate buffer Substances 0.000 description 2
- 238000000326 densiometry Methods 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000011859 microparticle Substances 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 150000007523 nucleic acids Chemical group 0.000 description 2
- 239000007764 o/w emulsion Substances 0.000 description 2
- IOQPZZOEVPZRBK-UHFFFAOYSA-N octan-1-amine Chemical compound CCCCCCCCN IOQPZZOEVPZRBK-UHFFFAOYSA-N 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 239000012466 permeate Substances 0.000 description 2
- 150000004713 phosphodiesters Chemical class 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 239000008057 potassium phosphate buffer Substances 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 230000002685 pulmonary effect Effects 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000002336 ribonucleotide Substances 0.000 description 2
- 125000002652 ribonucleotide group Chemical group 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 159000000000 sodium salts Chemical class 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000009885 systemic effect Effects 0.000 description 2
- 210000002845 virion Anatomy 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000001262 western blot Methods 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
- LRSASMSXMSNRBT-UHFFFAOYSA-N 5-methylcytosine Chemical group CC1=CNC(=O)N=C1N LRSASMSXMSNRBT-UHFFFAOYSA-N 0.000 description 1
- 102100037435 Antiviral innate immune response receptor RIG-I Human genes 0.000 description 1
- 101710127675 Antiviral innate immune response receptor RIG-I Proteins 0.000 description 1
- 241000180579 Arca Species 0.000 description 1
- FTEDXVNDVHYDQW-UHFFFAOYSA-N BAPTA Chemical compound OC(=O)CN(CC(O)=O)C1=CC=CC=C1OCCOC1=CC=CC=C1N(CC(O)=O)CC(O)=O FTEDXVNDVHYDQW-UHFFFAOYSA-N 0.000 description 1
- 108091032955 Bacterial small RNA Proteins 0.000 description 1
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 241000282994 Cervidae Species 0.000 description 1
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 1
- 208000017667 Chronic Disease Diseases 0.000 description 1
- 238000012286 ELISA Assay Methods 0.000 description 1
- 241000710945 Eastern equine encephalitis virus Species 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 108010068370 Glutens Proteins 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 description 1
- 101800000515 Non-structural protein 3 Proteins 0.000 description 1
- RSPISYXLHRIGJD-UHFFFAOYSA-N OOOO Chemical compound OOOO RSPISYXLHRIGJD-UHFFFAOYSA-N 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 229920012266 Poly(ether sulfone) PES Polymers 0.000 description 1
- 102000015623 Polynucleotide Adenylyltransferase Human genes 0.000 description 1
- 108010024055 Polynucleotide adenylyltransferase Proteins 0.000 description 1
- 108010076039 Polyproteins Proteins 0.000 description 1
- 241000206607 Porphyra umbilicalis Species 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 101800000980 Protease nsP2 Proteins 0.000 description 1
- 229930185560 Pseudouridine Natural products 0.000 description 1
- PTJWIQPHWPFNBW-UHFFFAOYSA-N Pseudouridine C Natural products OC1C(O)C(CO)OC1C1=CNC(=O)NC1=O PTJWIQPHWPFNBW-UHFFFAOYSA-N 0.000 description 1
- 230000006819 RNA synthesis Effects 0.000 description 1
- 230000004570 RNA-binding Effects 0.000 description 1
- 101800001758 RNA-directed RNA polymerase nsP4 Proteins 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 101710200092 Replicase polyprotein Proteins 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- 241000710961 Semliki Forest virus Species 0.000 description 1
- 241000710960 Sindbis virus Species 0.000 description 1
- 108020004459 Small interfering RNA Proteins 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 230000024932 T cell mediated immunity Effects 0.000 description 1
- 101710137500 T7 RNA polymerase Proteins 0.000 description 1
- 206010046865 Vaccinia virus infection Diseases 0.000 description 1
- 108020000999 Viral RNA Proteins 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000000240 adjuvant effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- FFBHFFJDDLITSX-UHFFFAOYSA-N benzyl N-[2-hydroxy-4-(3-oxomorpholin-4-yl)phenyl]carbamate Chemical compound OC1=C(NC(=O)OCC2=CC=CC=C2)C=CC(=C1)N1CCOCC1=O FFBHFFJDDLITSX-UHFFFAOYSA-N 0.000 description 1
- WGDUUQDYDIIBKT-UHFFFAOYSA-N beta-Pseudouridine Natural products OC1OC(CN2C=CC(=O)NC2=O)C(O)C1O WGDUUQDYDIIBKT-UHFFFAOYSA-N 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 150000001860 citric acid derivatives Chemical class 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000002681 effect on RNA Effects 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- DEFVIWRASFVYLL-UHFFFAOYSA-N ethylene glycol bis(2-aminoethyl)tetraacetic acid Chemical compound OC(=O)CN(CC(O)=O)CCOCCOCCN(CC(O)=O)CC(O)=O DEFVIWRASFVYLL-UHFFFAOYSA-N 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000005194 fractionation Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 235000021312 gluten Nutrition 0.000 description 1
- 238000011194 good manufacturing practice Methods 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000000091 immunopotentiator Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 238000010829 isocratic elution Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 102100039604 mRNA guanylyltransferase Human genes 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000009285 membrane fouling Methods 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- YACKEPLHDIMKIO-UHFFFAOYSA-N methylphosphonic acid Chemical compound CP(O)(O)=O YACKEPLHDIMKIO-UHFFFAOYSA-N 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 238000000424 optical density measurement Methods 0.000 description 1
- 229960003330 pentetic acid Drugs 0.000 description 1
- 229960005323 phenoxyethanol Drugs 0.000 description 1
- PTMHPRAIXMAOOB-UHFFFAOYSA-L phosphoramidate Chemical compound NP([O-])([O-])=O PTMHPRAIXMAOOB-UHFFFAOYSA-L 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 159000000001 potassium salts Chemical class 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000004845 protein aggregation Effects 0.000 description 1
- PTJWIQPHWPFNBW-GBNDHIKLSA-N pseudouridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1C1=CNC(=O)NC1=O PTJWIQPHWPFNBW-GBNDHIKLSA-N 0.000 description 1
- 239000013014 purified material Substances 0.000 description 1
- 150000003230 pyrimidines Chemical class 0.000 description 1
- 230000001698 pyrogenic effect Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 239000010832 regulated medical waste Substances 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 125000000548 ribosyl group Chemical group C1([C@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 239000012723 sample buffer Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000008362 succinate buffer Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 description 1
- 229960004906 thiomersal Drugs 0.000 description 1
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 125000002264 triphosphate group Chemical group [H]OP(=O)(O[H])OP(=O)(O[H])OP(=O)(O[H])O* 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 208000007089 vaccinia Diseases 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C07H21/02—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with ribosyl as saccharide radical
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/36—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction, e.g. ion-exchange, ion-pair, ion-suppression or ion-exclusion
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 and B01D15/30 - B01D15/36, e.g. affinity, ligand exchange or chiral chromatography
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 and B01D15/30 - B01D15/36, e.g. affinity, ligand exchange or chiral chromatography
- B01D15/3847—Multimodal interactions
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D61/00—Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor
- B01D61/14—Ultrafiltration; Microfiltration
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
- C07H21/04—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
-
- 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
- C12N15/101—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 by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 and B01D15/30 - B01D15/36, e.g. affinity, ligand exchange or chiral chromatography
- B01D15/3804—Affinity chromatography
- B01D15/3828—Ligand exchange chromatography, e.g. complexation, chelation or metal interaction chromatography
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Water Supply & Treatment (AREA)
- Crystallography & Structural Chemistry (AREA)
- Immunology (AREA)
- Veterinary Medicine (AREA)
- General Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Saccharide Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Claims (13)
1. Fremgangsmåde til oprensning af RNA fra en in w'fro-transskription-(IVT)-reaktionsprøve omfattende et første trin af core-bead-flow-through-kromatografi under tilstedeværelse af en buffer indeholdende et kaliumsalt, hvor fremgangsmåden yderligere omfatter, i et eller flere trin, udskiftning af buffer omfattende tangentialstrømsfiltrering.
2. Fremgangsmåde ifølge krav 1, hvor kaliumsaltet er kaliumfosfat eller kaliumklorid.
3. Fremgangsmåde ifølge krav 2, hvor kaliumsaltet er kaliumklorid i en koncentration på 100-500 mM.
4. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor RNA'et oprenses i præparativ skala.
5. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor fremgangsmåden ikke omfatter anvendelsen af litiumklorid, organiske opløsningsmidler, temperaturer højere end 70°C og/eller enzymatisk nedbrydning af DNA.
6. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor fremgangsmåden indbefatter kassering af materialer, der ikke indeholder RNA eller den ønskede slags RNA, og at bevare materialer, der indeholder RNA eller det ønskede RNA.
7. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor prøven indeholder RNA og et eller flere af: plasmid-DNA, deoxyoligonukleotider, deoxynucleosidmonofosfater, ribonucleosidtrifosfater og protein.
8. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor prøven indeholder RNA og ikke mere end fire af: plasmid-DNA, deoxyoligonukleotider, deoxynucleosidmonofosfater, ribonucleosidtrifosfater og protein; og eventuelt hvor prøven ikke indeholder genomisk DNA og/eller en cellemembran eller fragmenter deraf.
9. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor RNA'et er et enkeltstrenget RNA, som for eksempel et mRNA.
10. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor RNA'et omfatter en lineær sekvens på mindst 1.000 nukleotider, f.eks. mindst 5.000 nukleotider.
11. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor fremgangsmåden anvender en hydrofil stationær fase og/eller en hydrofil membran.
12. Fremgangsmåde ifølge et hvilket som helst af de foregående krav, hvor fremgangsmåden yderligere omfatter et trin med RNA-fremstilling, for eksempel ved anvendelse af in v/'tro-transskription af RNA.
13. Fremgangsmåde til fremstilling af en farmaceutisk sammensætning omfattende trinnene: (a) at oprense RNA ifølge en fremgangsmåde ifølge et hvilket som helst af kravene 1-12; og (b) at formulere det oprensede RNA som en farmaceutisk sammensætning.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201361799705P | 2013-03-15 | 2013-03-15 | |
| PCT/EP2014/055014 WO2014140211A1 (en) | 2013-03-15 | 2014-03-13 | Rna purification methods |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DK2970948T3 true DK2970948T3 (da) | 2019-04-08 |
Family
ID=50272650
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DK14709704.2T DK2970948T3 (da) | 2013-03-15 | 2014-03-13 | Fremgangsmåder til rna-oprensning |
Country Status (17)
| Country | Link |
|---|---|
| US (3) | US11155572B2 (da) |
| EP (2) | EP2970948B1 (da) |
| JP (1) | JP6461830B2 (da) |
| CN (1) | CN105283548A (da) |
| BR (1) | BR112015023513A2 (da) |
| CA (1) | CA2906281A1 (da) |
| CY (1) | CY1121478T1 (da) |
| DK (1) | DK2970948T3 (da) |
| ES (2) | ES2923018T3 (da) |
| HR (1) | HRP20190197T1 (da) |
| HU (1) | HUE042072T2 (da) |
| LT (1) | LT2970948T (da) |
| PL (1) | PL2970948T3 (da) |
| PT (1) | PT2970948T (da) |
| SI (1) | SI2970948T1 (da) |
| TR (1) | TR201903248T4 (da) |
| WO (1) | WO2014140211A1 (da) |
Families Citing this family (53)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US9464124B2 (en) | 2011-09-12 | 2016-10-11 | Moderna Therapeutics, Inc. | Engineered nucleic acids and methods of use thereof |
| JP6586075B2 (ja) * | 2013-03-14 | 2019-10-02 | トランスレイト バイオ， インコーポレイテッド | メッセンジャーｒｎａの精製方法 |
| WO2014152031A1 (en) | 2013-03-15 | 2014-09-25 | Moderna Therapeutics, Inc. | Ribonucleic acid purification |
| DK2970948T3 (da) * | 2013-03-15 | 2019-04-08 | Glaxosmithkline Biologicals Sa | Fremgangsmåder til rna-oprensning |
| WO2015048744A2 (en) | 2013-09-30 | 2015-04-02 | Moderna Therapeutics, Inc. | Polynucleotides encoding immune modulating polypeptides |
| EP3052521A1 (en) | 2013-10-03 | 2016-08-10 | Moderna Therapeutics, Inc. | Polynucleotides encoding low density lipoprotein receptor |
| PL4023249T3 (pl) | 2014-04-23 | 2025-03-10 | Modernatx, Inc. | Szczepionki z kwasem nukleinowym |
| US9850269B2 (en) | 2014-04-25 | 2017-12-26 | Translate Bio, Inc. | Methods for purification of messenger RNA |
| WO2016180430A1 (en) | 2015-05-08 | 2016-11-17 | Curevac Ag | Method for producing rna |
| EP4660315A3 (en) | 2015-05-29 | 2026-03-18 | CureVac Manufacturing GmbH | A method for producing and purifying rna, comprising at least one step of tangential flow filtration |
| WO2017025120A1 (en) | 2015-08-07 | 2017-02-16 | Curevac Ag | Process for the in vivo production of rna in a host cell |
| WO2017083286A1 (en) | 2015-11-09 | 2017-05-18 | University Of Notre Dame | Particle size purification method and devices |
| MX2018007987A (es) | 2015-12-28 | 2019-01-10 | Novartis Ag | Composiciones y metodos para el tratamiento de hemoglobinopatias. |
| WO2017223176A1 (en) | 2016-06-24 | 2017-12-28 | Modernatx, Inc. | Methods and apparatus for filtration |
| IL301963A (en) * | 2016-07-21 | 2023-06-01 | Spark Therapeutics Inc | Scalable High Recovery Methods for High-Throughput Recombinant Adeno-Associated Viral Vector Production and Recombinant Adeno-Associated Viral Vectors Made So |
| WO2018041921A1 (en) | 2016-08-31 | 2018-03-08 | Curevac Ag | Mixing device for the production of a liquid nucleic acid composition |
| CA3036831A1 (en) | 2016-09-14 | 2018-03-22 | Modernatx, Inc. | High purity rna compositions and methods for preparation thereof |
| WO2018096179A1 (en) | 2016-11-28 | 2018-05-31 | Curevac Ag | Method for purifying rna |
| US11384352B2 (en) | 2016-12-13 | 2022-07-12 | Modernatx, Inc. | RNA affinity purification |
| TW201839136A (zh) | 2017-02-06 | 2018-11-01 | 瑞士商諾華公司 | 治療血色素異常症之組合物及方法 |
| CA3054321A1 (en) * | 2017-02-27 | 2018-08-30 | Translate Bio, Inc. | Methods for purification of messenger rna |
| EP3625363A1 (en) | 2017-05-17 | 2020-03-25 | CureVac Real Estate GmbH | Method for determining at least one quality parameter of an rna sample |
| MA49421A (fr) | 2017-06-15 | 2020-04-22 | Modernatx Inc | Formulations d'arn |
| US12215378B2 (en) | 2017-08-11 | 2025-02-04 | Glaxosmithkline Biologicals Sa | RNA identity method using RNAse H digestion and size fractionating |
| EP3668979A4 (en) | 2017-08-18 | 2021-06-02 | Modernatx, Inc. | METHOD OF HPLC ANALYSIS |
| MA49914A (fr) | 2017-08-18 | 2021-04-21 | Modernatx Inc | Procédés analytiques par hplc |
| EP3675817A1 (en) | 2017-08-31 | 2020-07-08 | Modernatx, Inc. | Methods of making lipid nanoparticles |
| TWI816721B (zh) * | 2017-12-07 | 2023-10-01 | 德商Ｅｍｐ生物科技公司 | 應用徑向技術色層分析術的系統及方法 |
| WO2020002598A1 (en) * | 2018-06-28 | 2020-01-02 | Curevac Ag | Bioreactor for rna in vitro transcription |
| KR102950141B1 (ko) | 2018-08-24 | 2026-04-07 | 트랜슬레이트 바이오 인코포레이티드 | 전령 rna의 정제 방법 |
| WO2020061457A1 (en) | 2018-09-20 | 2020-03-26 | Modernatx, Inc. | Preparation of lipid nanoparticles and methods of administration thereof |
| IL282368B2 (en) * | 2018-10-24 | 2024-09-01 | Hoffmann La Roche | Process for the purification of oligonucleotides |
| WO2020160087A1 (en) * | 2019-01-29 | 2020-08-06 | Flagship Pioneering Innovations V, Inc. | Methods of separating long polynucleotides from a composition |
| AU2020220357B2 (en) * | 2019-02-11 | 2025-06-26 | Ethris Gmbh | mRNA purification by tangential flow filtration |
| CN113874502A (zh) | 2019-03-11 | 2021-12-31 | 摩登纳特斯有限公司 | 补料分批体外转录方法 |
| US20220306678A1 (en) * | 2019-06-20 | 2022-09-29 | Arcturus Therapeutics, Inc. | Improved in vitro transcription purification platform |
| JP7846007B2 (ja) | 2019-12-18 | 2026-04-14 | ノバルティス アーゲー | 異常ヘモグロビン症の治療用組成物及び方法 |
| US12194089B2 (en) | 2020-02-04 | 2025-01-14 | CureVac SE | Coronavirus vaccine |
| JP2023515485A (ja) * | 2020-02-21 | 2023-04-13 | バイオジェン・エムエイ・インコーポレイテッド | 限外濾過／透析濾過を用いたオリゴヌクレオチド組成物の調製方法 |
| US20230265476A1 (en) * | 2020-05-12 | 2023-08-24 | Charalampos Makatsoris | A method and modular apparatus for the synthesis of rna-based therapeutics |
| US12188060B2 (en) | 2020-05-15 | 2025-01-07 | Crispr Therapeutics Ag | Messenger RNA encoding Cas9 for use in genome-editing systems |
| CN115715324A (zh) * | 2020-06-19 | 2023-02-24 | 伊泽阿恩埃免疫疗法股份有限公司 | Rna纯化方法 |
| CN111812230A (zh) * | 2020-06-24 | 2020-10-23 | 南京济群生物科技有限公司 | 一种半定量测定dna中rna含量的sec-hplc检测方法 |
| JP7657918B2 (ja) * | 2020-09-14 | 2025-04-07 | エンクイスト テクノロジーズ インコーポレイテッド | 生物学的生成物を精製する方法 |
| EP4274607A1 (en) | 2021-01-11 | 2023-11-15 | ModernaTX, Inc. | Seasonal rna influenza virus vaccines |
| US20220364078A1 (en) * | 2021-05-14 | 2022-11-17 | Crispr Therapeutics Ag | Mrna large scale synthesis and purification |
| US20240218353A1 (en) * | 2021-06-17 | 2024-07-04 | Modernatx, Inc. | Alternative rna purification strategies |
| US20250127809A1 (en) | 2021-06-23 | 2025-04-24 | Novartis Ag | Compositions and methods for the treatment of hemoglobinopathies |
| US20250137024A1 (en) | 2021-09-01 | 2025-05-01 | Glaxosmithkline Biologicals Sa | Plasmid dna purification methods |
| EP4422698A1 (en) | 2021-10-29 | 2024-09-04 | CureVac SE | Improved circular rna for expressing therapeutic proteins |
| CN121046364A (zh) | 2022-07-29 | 2025-12-02 | 赛诺菲巴斯德有限公司 | 用于无乙醇mRNA纯化的方法 |
| WO2024072904A2 (en) * | 2022-09-28 | 2024-04-04 | Modernatx, Inc. | Purification of nucleotides |
| WO2025186218A1 (en) | 2024-03-04 | 2025-09-12 | Cytiva Bioprocess R&D Ab | Purification and analysis of rna |
Family Cites Families (26)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5256294A (en) | 1990-09-17 | 1993-10-26 | Genentech, Inc. | Tangential flow filtration process and apparatus |
| US6011148A (en) | 1996-08-01 | 2000-01-04 | Megabios Corporation | Methods for purifying nucleic acids |
| EP0853123A1 (de) | 1997-01-10 | 1998-07-15 | Roche Diagnostics GmbH | Reinigung von DNA durch Cross-Flow-Filtration |
| WO1999040098A1 (de) | 1998-02-04 | 1999-08-12 | Merck Patent Gmbh | Verfahren zur isolierung und aufreinigung von nucleinsäuren |
| EP1488233A4 (en) * | 2001-10-09 | 2006-06-21 | Genentech Inc | NEW ACID MUDE PROTEINS AND THESE CODING POLYNUCLEOTIDES |
| US8075780B2 (en) | 2003-11-24 | 2011-12-13 | Millipore Corporation | Purification and concentration of synthetic biological molecules |
| CN1989250B (zh) | 2004-05-18 | 2013-11-20 | 阿尔法瓦克斯公司 | Tc-83衍生的甲病毒属载体、颗粒和方法 |
| US20080207487A1 (en) * | 2006-11-02 | 2008-08-28 | Neose Technologies, Inc. | Manufacturing process for the production of polypeptides expressed in insect cell-lines |
| DE102006061015A1 (de) | 2006-12-22 | 2008-06-26 | Curevac Gmbh | Verfahren zur Reinigung von RNA im präparativen Maßstab mittels HPLC |
| CA2766907A1 (en) | 2009-07-06 | 2011-01-13 | Novartis Ag | Self replicating rna molecules and uses thereof |
| EP2591114B1 (en) | 2010-07-06 | 2016-06-08 | GlaxoSmithKline Biologicals SA | Immunisation of large mammals with low doses of rna |
| MX342608B (es) | 2010-07-06 | 2016-10-06 | Novartis Ag * | Particulas de suministro similares a viriones para moleculas de arn de autorreplicacion. |
| US9192661B2 (en) | 2010-07-06 | 2015-11-24 | Novartis Ag | Delivery of self-replicating RNA using biodegradable polymer particles |
| NZ606591A (en) | 2010-07-06 | 2015-02-27 | Novartis Ag | Cationic oil-in-water emulsions |
| WO2012006732A1 (en) * | 2010-07-16 | 2012-01-19 | Universite De Montreal | Affinity purification of rna under native conditions based on the lambda boxb/n peptide interaction |
| LT4226941T (lt) | 2010-08-31 | 2025-01-10 | Glaxosmithkline Biologicals Sa | Pegilintos liposomos, skirtos imunogeną koduojančios rnr pristatymui |
| SI4043040T1 (sl) | 2010-08-31 | 2023-04-28 | Glaxosmithkline Biologicals Sa | Mali liposomi za dostavo imunogen-kodirajoče RNA |
| MX2013002332A (es) | 2010-08-31 | 2013-03-18 | Novartis Ag | Lipidos apropiados para suministro liposomal del arn que codifica la proteina. |
| CA2820861A1 (en) | 2010-12-10 | 2012-06-14 | Tracy Thompson | Compositions for separation methods |
| MX350258B (es) | 2011-07-06 | 2017-08-31 | Novartis Ag | Emulsiones cationicas de aceite en agua. |
| EP2729165B1 (en) | 2011-07-06 | 2017-11-08 | GlaxoSmithKline Biologicals SA | Immunogenic combination compositions and uses thereof |
| JP6059220B2 (ja) | 2011-07-06 | 2017-01-18 | ノバルティス アーゲー | 核酸を含む水中油型エマルジョン |
| EP4115875A1 (en) | 2011-07-06 | 2023-01-11 | GlaxoSmithKline Biologicals S.A. | Liposomes having useful n:p ratio for delivery of rna molecules |
| WO2013103439A1 (en) * | 2012-01-06 | 2013-07-11 | Infinity Fuel Cell And Hydrogen, Inc. | Electrochemical cell with improved water or gas management |
| JP6586075B2 (ja) | 2013-03-14 | 2019-10-02 | トランスレイト バイオ， インコーポレイテッド | メッセンジャーｒｎａの精製方法 |
| DK2970948T3 (da) * | 2013-03-15 | 2019-04-08 | Glaxosmithkline Biologicals Sa | Fremgangsmåder til rna-oprensning |
-
2014
- 2014-03-13 DK DK14709704.2T patent/DK2970948T3/da active
- 2014-03-13 SI SI201431108T patent/SI2970948T1/sl unknown
- 2014-03-13 ES ES18212923T patent/ES2923018T3/es active Active
- 2014-03-13 BR BR112015023513A patent/BR112015023513A2/pt not_active Application Discontinuation
- 2014-03-13 JP JP2015562160A patent/JP6461830B2/ja active Active
- 2014-03-13 US US14/775,760 patent/US11155572B2/en active Active
- 2014-03-13 PL PL14709704T patent/PL2970948T3/pl unknown
- 2014-03-13 HU HUE14709704A patent/HUE042072T2/hu unknown
- 2014-03-13 PT PT14709704T patent/PT2970948T/pt unknown
- 2014-03-13 CN CN201480015831.4A patent/CN105283548A/zh active Pending
- 2014-03-13 ES ES14709704T patent/ES2720200T3/es active Active
- 2014-03-13 EP EP14709704.2A patent/EP2970948B1/en active Active
- 2014-03-13 WO PCT/EP2014/055014 patent/WO2014140211A1/en not_active Ceased
- 2014-03-13 LT LTEP14709704.2T patent/LT2970948T/lt unknown
- 2014-03-13 HR HRP20190197TT patent/HRP20190197T1/hr unknown
- 2014-03-13 TR TR2019/03248T patent/TR201903248T4/tr unknown
- 2014-03-13 EP EP18212923.9A patent/EP3521429B1/en active Active
- 2014-03-13 CA CA2906281A patent/CA2906281A1/en active Pending
-
2019
- 2019-03-12 CY CY20191100284T patent/CY1121478T1/el unknown
-
2021
- 2021-01-25 US US17/157,150 patent/US12018044B2/en active Active
-
2024
- 2024-05-15 US US18/664,453 patent/US20240300994A1/en active Pending
Also Published As
| Publication number | Publication date |
|---|---|
| CN105283548A (zh) | 2016-01-27 |
| EP3521429B1 (en) | 2022-05-04 |
| HRP20190197T1 (hr) | 2019-04-05 |
| JP2016512212A (ja) | 2016-04-25 |
| PT2970948T (pt) | 2019-03-20 |
| PL2970948T3 (pl) | 2019-06-28 |
| ES2720200T3 (es) | 2019-07-18 |
| CY1121478T1 (el) | 2020-05-29 |
| EP3521429A1 (en) | 2019-08-07 |
| US20210214388A1 (en) | 2021-07-15 |
| EP2970948A1 (en) | 2016-01-20 |
| TR201903248T4 (tr) | 2019-03-21 |
| ES2923018T3 (es) | 2022-09-22 |
| US20160024139A1 (en) | 2016-01-28 |
| US12018044B2 (en) | 2024-06-25 |
| LT2970948T (lt) | 2019-03-25 |
| US11155572B2 (en) | 2021-10-26 |
| EP2970948B1 (en) | 2018-12-26 |
| WO2014140211A1 (en) | 2014-09-18 |
| BR112015023513A2 (pt) | 2017-07-18 |
| JP6461830B2 (ja) | 2019-01-30 |
| HUE042072T2 (hu) | 2019-06-28 |
| SI2970948T1 (sl) | 2019-04-30 |
| US20240300994A1 (en) | 2024-09-12 |
| CA2906281A1 (en) | 2014-09-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20240300994A1 (en) | Rna purification methods | |
| JP5129805B2 (ja) | 細胞培養物から精製水疱性口内炎ウイルスを単離するための精製プロセス | |
| JP7668231B2 (ja) | メッセンジャーｒｎａの精製方法 | |
| DE202016009003U1 (de) | Zusammensetzung umfassend in vitro transkribierte RNA erhältlich durch ein Verfahren zur Herstellung und Reinigung von RNA mit mindestens einem Schritt mit einer tangentialen Flussfiltration | |
| Steppert et al. | A scalable, integrated downstream process for production of a recombinant measles virus-vectored vaccine | |
| EP3581646A1 (en) | Integrated manufacturing and chromatographic system for virus production | |
| CN106574252B (zh) | 从细胞培养物中纯化脊髓灰质炎病毒的方法 | |
| US20150030565A1 (en) | Purification of flaviviruses | |
| US20250101409A1 (en) | Methods for ethanol-free mrna purification | |
| Cui et al. | Comprehensive studies on building a scalable downstream process for mRNAs to enable mRNA therapeutics | |
| B. Carvalho et al. | Downstream processing for influenza vaccines and candidates: An update | |
| EP3371302B1 (en) | Method for the separation of virus compositions including depletion and purification thereof | |
| JP5614936B2 (ja) | アニオン交換基が固定された多孔膜を用いた核酸の精製方法 | |
| EP3921428B1 (en) | Methods for providing purified viral particles of semliki forest virus (sfv), preparations obtainable thereby, and uses thereof | |
| Mundle et al. | Preparation of pure, high titer, pseudoinfectious Flavivirus particles by hollow fiber tangential flow filtration and anion exchange chromatography | |
| WO2025163146A1 (en) | Methods for ethanol-free mrna purification | |
| WO2024131232A1 (zh) | 环状rna分离和纯化方法 | |
| EP3165604A1 (en) | Method for the purification of virus compositions and virus compositions obtained |