US20080154029A1 - Process For Manufacturing a Composite Sorbent Material For Chromatographical Separation of Biopolymers - Google Patents
Process For Manufacturing a Composite Sorbent Material For Chromatographical Separation of Biopolymers Download PDFInfo
- Publication number
- US20080154029A1 US20080154029A1 US11/547,011 US54701105A US2008154029A1 US 20080154029 A1 US20080154029 A1 US 20080154029A1 US 54701105 A US54701105 A US 54701105A US 2008154029 A1 US2008154029 A1 US 2008154029A1
- Authority
- US
- United States
- Prior art keywords
- sorbent material
- sorbent
- general formula
- composite
- support
- 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.)
- Abandoned
Links
- 239000002594 sorbent Substances 0.000 title claims abstract description 72
- 238000000926 separation method Methods 0.000 title claims abstract description 26
- 229920001222 biopolymer Polymers 0.000 title claims abstract description 12
- 239000002131 composite material Substances 0.000 title claims description 19
- 238000000034 method Methods 0.000 title claims description 18
- 238000004519 manufacturing process Methods 0.000 title claims description 3
- 239000000463 material Substances 0.000 claims abstract description 37
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 19
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 19
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 19
- 239000011248 coating agent Substances 0.000 claims abstract description 18
- 238000000576 coating method Methods 0.000 claims abstract description 18
- 229920001577 copolymer Polymers 0.000 claims abstract description 16
- 150000001875 compounds Chemical class 0.000 claims abstract description 9
- 239000000178 monomer Substances 0.000 claims abstract description 8
- 125000003118 aryl group Chemical group 0.000 claims abstract description 6
- DMZWVCJEOLBQCZ-UHFFFAOYSA-N chloro(ethenyl)silane Chemical compound Cl[SiH2]C=C DMZWVCJEOLBQCZ-UHFFFAOYSA-N 0.000 claims abstract description 6
- 238000005464 sample preparation method Methods 0.000 claims abstract description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 48
- 239000000872 buffer Substances 0.000 claims description 37
- 239000011148 porous material Substances 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- -1 hydroxylalkyl ester Chemical class 0.000 claims description 22
- 239000000203 mixture Substances 0.000 claims description 21
- 229920000642 polymer Polymers 0.000 claims description 17
- 102000004169 proteins and genes Human genes 0.000 claims description 16
- 108090000623 proteins and genes Proteins 0.000 claims description 14
- 239000000126 substance Substances 0.000 claims description 12
- MYRTYDVEIRVNKP-UHFFFAOYSA-N divinylbenzene Substances C=CC1=CC=CC=C1C=C MYRTYDVEIRVNKP-UHFFFAOYSA-N 0.000 claims description 11
- 150000003839 salts Chemical class 0.000 claims description 10
- 238000009826 distribution Methods 0.000 claims description 9
- PPBRXRYQALVLMV-UHFFFAOYSA-N Styrene Natural products C=CC1=CC=CC=C1 PPBRXRYQALVLMV-UHFFFAOYSA-N 0.000 claims description 7
- 238000013375 chromatographic separation Methods 0.000 claims description 7
- 230000014759 maintenance of location Effects 0.000 claims description 7
- 229920006395 saturated elastomer Polymers 0.000 claims description 7
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- 150000007824 aliphatic compounds Chemical class 0.000 claims description 6
- 125000000217 alkyl group Chemical group 0.000 claims description 6
- 150000001491 aromatic compounds Chemical class 0.000 claims description 6
- 230000000379 polymerizing effect Effects 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 5
- 239000011159 matrix material Substances 0.000 claims description 5
- 230000000274 adsorptive effect Effects 0.000 claims description 4
- 150000001298 alcohols Chemical class 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 239000003960 organic solvent Substances 0.000 claims description 4
- IGGDKDTUCAWDAN-UHFFFAOYSA-N 1-vinylnaphthalene Chemical class C1=CC=C2C(C=C)=CC=CC2=C1 IGGDKDTUCAWDAN-UHFFFAOYSA-N 0.000 claims description 3
- 150000001735 carboxylic acids Chemical class 0.000 claims description 3
- 239000005289 controlled pore glass Substances 0.000 claims description 3
- 239000003995 emulsifying agent Substances 0.000 claims description 3
- 125000005842 heteroatom Chemical group 0.000 claims description 3
- 229910010272 inorganic material Inorganic materials 0.000 claims description 3
- 239000011147 inorganic material Substances 0.000 claims description 3
- 229910044991 metal oxide Inorganic materials 0.000 claims description 3
- 150000004706 metal oxides Chemical class 0.000 claims description 3
- 230000000717 retained effect Effects 0.000 claims description 3
- 150000003440 styrenes Chemical class 0.000 claims description 3
- 108091005461 Nucleic proteins Proteins 0.000 claims description 2
- RTAQQCXQSZGOHL-UHFFFAOYSA-N Titanium Chemical compound [Ti] RTAQQCXQSZGOHL-UHFFFAOYSA-N 0.000 claims description 2
- QCWXUUIWCKQGHC-UHFFFAOYSA-N Zirconium Chemical compound [Zr] QCWXUUIWCKQGHC-UHFFFAOYSA-N 0.000 claims description 2
- 229910052782 aluminium Inorganic materials 0.000 claims description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 2
- 238000006116 polymerization reaction Methods 0.000 claims description 2
- 239000010936 titanium Substances 0.000 claims description 2
- 229910052719 titanium Inorganic materials 0.000 claims description 2
- 229910052726 zirconium Inorganic materials 0.000 claims description 2
- 239000005909 Kieselgur Substances 0.000 claims 1
- 238000007720 emulsion polymerization reaction Methods 0.000 claims 1
- UQSXHKLRYXJYBZ-UHFFFAOYSA-N iron oxide Inorganic materials [Fe]=O UQSXHKLRYXJYBZ-UHFFFAOYSA-N 0.000 claims 1
- 235000013980 iron oxide Nutrition 0.000 claims 1
- VBMVTYDPPZVILR-UHFFFAOYSA-N iron(2+);oxygen(2-) Chemical class [O-2].[Fe+2] VBMVTYDPPZVILR-UHFFFAOYSA-N 0.000 claims 1
- 239000003505 polymerization initiator Substances 0.000 claims 1
- LIVNPJMFVYWSIS-UHFFFAOYSA-N silicon monoxide Chemical class [Si-]#[O+] LIVNPJMFVYWSIS-UHFFFAOYSA-N 0.000 claims 1
- 229910052814 silicon oxide Inorganic materials 0.000 claims 1
- 238000002955 isolation Methods 0.000 abstract description 18
- 239000006166 lysate Substances 0.000 abstract description 13
- 238000000746 purification Methods 0.000 abstract description 12
- 239000007787 solid Substances 0.000 abstract description 6
- 238000004132 cross linking Methods 0.000 abstract description 3
- 150000002148 esters Chemical class 0.000 abstract description 3
- 150000002170 ethers Chemical class 0.000 abstract description 3
- 108020004414 DNA Proteins 0.000 description 37
- 239000000243 solution Substances 0.000 description 19
- 239000000377 silicon dioxide Substances 0.000 description 16
- 0 CB*CC(C(C)C)[Si](C)(C)OC Chemical compound CB*CC(C(C)C)[Si](C)(C)OC 0.000 description 15
- 239000000741 silica gel Substances 0.000 description 14
- 229910002027 silica gel Inorganic materials 0.000 description 14
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 9
- 230000009089 cytolysis Effects 0.000 description 9
- 238000002360 preparation method Methods 0.000 description 9
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 8
- 229940098773 bovine serum albumin Drugs 0.000 description 8
- 229910000366 copper(II) sulfate Inorganic materials 0.000 description 8
- 229910052681 coesite Inorganic materials 0.000 description 7
- 229910052906 cristobalite Inorganic materials 0.000 description 7
- 229910052682 stishovite Inorganic materials 0.000 description 7
- 229910052905 tridymite Inorganic materials 0.000 description 7
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 6
- 102000016943 Muramidase Human genes 0.000 description 6
- 108010014251 Muramidase Proteins 0.000 description 6
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 6
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 6
- 239000004325 lysozyme Substances 0.000 description 6
- 229960000274 lysozyme Drugs 0.000 description 6
- 235000010335 lysozyme Nutrition 0.000 description 6
- 239000002245 particle Substances 0.000 description 6
- 150000003384 small molecules Chemical class 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- XXROGKLTLUQVRX-UHFFFAOYSA-N allyl alcohol Chemical compound OCC=C XXROGKLTLUQVRX-UHFFFAOYSA-N 0.000 description 5
- 238000009835 boiling Methods 0.000 description 5
- 229940116441 divinylbenzene Drugs 0.000 description 5
- 230000002209 hydrophobic effect Effects 0.000 description 5
- 239000011347 resin Substances 0.000 description 5
- 229920005989 resin Polymers 0.000 description 5
- 238000001179 sorption measurement Methods 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 4
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 102000006382 Ribonucleases Human genes 0.000 description 4
- 108010083644 Ribonucleases Proteins 0.000 description 4
- XSDCTSITJJJDPY-UHFFFAOYSA-N chloro-ethenyl-dimethylsilane Chemical compound C[Si](C)(Cl)C=C XSDCTSITJJJDPY-UHFFFAOYSA-N 0.000 description 4
- 238000001704 evaporation Methods 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 239000003999 initiator Substances 0.000 description 4
- 230000010874 maintenance of protein location Effects 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 230000008020 evaporation Effects 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 239000010410 layer Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000003260 vortexing Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- BLIQUJLAJXRXSG-UHFFFAOYSA-N 1-benzyl-3-(trifluoromethyl)pyrrolidin-1-ium-3-carboxylate Chemical compound C1C(C(=O)O)(C(F)(F)F)CCN1CC1=CC=CC=C1 BLIQUJLAJXRXSG-UHFFFAOYSA-N 0.000 description 2
- JESXATFQYMPTNL-UHFFFAOYSA-N 2-ethenylphenol Chemical compound OC1=CC=CC=C1C=C JESXATFQYMPTNL-UHFFFAOYSA-N 0.000 description 2
- 229940044192 2-hydroxyethyl methacrylate Drugs 0.000 description 2
- 239000004342 Benzoyl peroxide Substances 0.000 description 2
- OMPJBNCRMGITSC-UHFFFAOYSA-N Benzoylperoxide Chemical compound C=1C=CC=CC=1C(=O)OOC(=O)C1=CC=CC=C1 OMPJBNCRMGITSC-UHFFFAOYSA-N 0.000 description 2
- WDWDHNJBYJURRG-IHWYPQMZSA-N C/C(/N)=C(\C(ON)=O)/N Chemical compound C/C(/N)=C(\C(ON)=O)/N WDWDHNJBYJURRG-IHWYPQMZSA-N 0.000 description 2
- JZXCMFPZDQXSDM-UHFFFAOYSA-N CCCCC(C(C)C)[Si](C)(C)OC Chemical compound CCCCC(C(C)C)[Si](C)(C)OC JZXCMFPZDQXSDM-UHFFFAOYSA-N 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 230000005526 G1 to G0 transition Effects 0.000 description 2
- WOBHKFSMXKNTIM-UHFFFAOYSA-N Hydroxyethyl methacrylate Chemical compound CC(=C)C(=O)OCCO WOBHKFSMXKNTIM-UHFFFAOYSA-N 0.000 description 2
- 208000029422 Hypernatremia Diseases 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- CERQOIWHTDAKMF-UHFFFAOYSA-N Methacrylic acid Chemical compound CC(=C)C(O)=O CERQOIWHTDAKMF-UHFFFAOYSA-N 0.000 description 2
- 125000003545 alkoxy group Chemical group 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- UPBRSCSZHBHGHS-UHFFFAOYSA-N benzene 1,2-bis(ethenyl)benzene Chemical compound C1=CC=CC=C1.C=CC1=CC=CC=C1C=C UPBRSCSZHBHGHS-UHFFFAOYSA-N 0.000 description 2
- 235000019400 benzoyl peroxide Nutrition 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000000470 constituent Substances 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 125000004663 dialkyl amino group Chemical group 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- FJKIXWOMBXYWOQ-UHFFFAOYSA-N ethenoxyethane Chemical compound CCOC=C FJKIXWOMBXYWOQ-UHFFFAOYSA-N 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 238000004191 hydrophobic interaction chromatography Methods 0.000 description 2
- 238000012417 linear regression Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 2
- 229910052753 mercury Inorganic materials 0.000 description 2
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 2
- 238000010992 reflux Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 229920002554 vinyl polymer Polymers 0.000 description 2
- QBDSMJHIEZXKKJ-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;2-hydroxyethyl 2-methylprop-2-enoate Chemical compound CC(=C)C(=O)OCCO.C=CC1=CC=CC=C1C=C QBDSMJHIEZXKKJ-UHFFFAOYSA-N 0.000 description 1
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 1
- ISRGONDNXBCDBM-UHFFFAOYSA-N 2-chlorostyrene Chemical compound ClC1=CC=CC=C1C=C ISRGONDNXBCDBM-UHFFFAOYSA-N 0.000 description 1
- 238000004438 BET method Methods 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 241000701959 Escherichia virus Lambda Species 0.000 description 1
- VGGSQFUCUMXWEO-UHFFFAOYSA-N Ethene Chemical compound C=C VGGSQFUCUMXWEO-UHFFFAOYSA-N 0.000 description 1
- 239000005977 Ethylene Substances 0.000 description 1
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 1
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 description 1
- 239000004677 Nylon Substances 0.000 description 1
- ZYFVNVRFVHJEIU-UHFFFAOYSA-N PicoGreen Chemical compound CN(C)CCCN(CCCN(C)C)C1=CC(=CC2=[N+](C3=CC=CC=C3S2)C)C2=CC=CC=C2N1C1=CC=CC=C1 ZYFVNVRFVHJEIU-UHFFFAOYSA-N 0.000 description 1
- 241001232115 Porogramme Species 0.000 description 1
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- ROOXNKNUYICQNP-UHFFFAOYSA-N ammonium peroxydisulfate Substances [NH4+].[NH4+].[O-]S(=O)(=O)OOS([O-])(=O)=O ROOXNKNUYICQNP-UHFFFAOYSA-N 0.000 description 1
- VAZSKTXWXKYQJF-UHFFFAOYSA-N ammonium persulfate Chemical compound [NH4+].[NH4+].[O-]S(=O)OOS([O-])=O VAZSKTXWXKYQJF-UHFFFAOYSA-N 0.000 description 1
- 229910001870 ammonium persulfate Inorganic materials 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000012159 carrier gas Substances 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000008119 colloidal silica Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000003297 denaturating effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 238000003682 fluorination reaction Methods 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000012456 homogeneous solution Substances 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000011261 inert gas Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 1
- 229920001778 nylon Polymers 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 239000013047 polymeric layer Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 229910000077 silane Inorganic materials 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- RYYKJJJTJZKILX-UHFFFAOYSA-M sodium octadecanoate Chemical compound [Na+].CCCCCCCCCCCCCCCCCC([O-])=O RYYKJJJTJZKILX-UHFFFAOYSA-M 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
Classifications
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/02—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material
- B01J20/10—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising silica or silicate
- B01J20/103—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising silica or silicate comprising silica
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
- B01J20/261—Synthetic macromolecular compounds obtained by reactions only involving carbon to carbon unsaturated bonds
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
- B01J20/265—Synthetic macromolecular compounds modified or post-treated polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28054—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their surface properties or porosity
- B01J20/28057—Surface area, e.g. B.E.T specific surface area
- B01J20/28059—Surface area, e.g. B.E.T specific surface area being less than 100 m2/g
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28054—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their surface properties or porosity
- B01J20/28057—Surface area, e.g. B.E.T specific surface area
- B01J20/28061—Surface area, e.g. B.E.T specific surface area being in the range 100-500 m2/g
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28054—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their surface properties or porosity
- B01J20/28078—Pore diameter
- B01J20/28083—Pore diameter being in the range 2-50 nm, i.e. mesopores
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28054—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their surface properties or porosity
- B01J20/28078—Pore diameter
- B01J20/28085—Pore diameter being more than 50 nm, i.e. macropores
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
- B01J20/28054—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their surface properties or porosity
- B01J20/28088—Pore-size distribution
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/282—Porous sorbents
- B01J20/283—Porous sorbents based on silica
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to polymers
- B01J20/289—Phases chemically bonded to a substrate, e.g. to silica or to polymers bonded via a spacer
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3202—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the carrier, support or substrate used for impregnation or coating
- B01J20/3204—Inorganic carriers, supports or substrates
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3214—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the method for obtaining this coating or impregnating
- B01J20/3217—Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond
- B01J20/3219—Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond involving a particular spacer or linking group, e.g. for attaching an active group
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
- B01J20/3268—Macromolecular compounds
- B01J20/3278—Polymers being grafted on the carrier
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F212/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an aromatic carbocyclic ring
- C08F212/02—Monomers containing only one unsaturated aliphatic radical
- C08F212/04—Monomers containing only one unsaturated aliphatic radical containing one ring
- C08F212/06—Hydrocarbons
- C08F212/08—Styrene
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F212/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by an aromatic carbocyclic ring
- C08F212/02—Monomers containing only one unsaturated aliphatic radical
- C08F212/04—Monomers containing only one unsaturated aliphatic radical containing one ring
- C08F212/14—Monomers containing only one unsaturated aliphatic radical containing one ring substituted by heteroatoms or groups containing heteroatoms
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F292/00—Macromolecular compounds obtained by polymerising monomers on to inorganic materials
Definitions
- the present invention relates to a sorbent material for separation and purification of biopolymers, particularly nucleic acids, having a solid support substantially modified with a copolymer coating comprising aromatic monomers and crosslinking compounds and unsaturated esters or ethers preferably attached to the support via a vinylchlorsilane.
- hydrophilic support material such as silica gel
- hydrophobic moieties like alkyl chains of different length.
- U.S. Pat. No. 4,045,353 describes a chromatographic material prepared by radiation of polymerisable monomers absorbed on a microporous inorganic substrate.
- the sorbent material thus obtained is suitable for separation of relatively small molecules, but there is insufficient selectivity for separation of biopolymers like nucleic acids.
- EP 1 148 945 discloses a material having a solid support of controlled pore glass and a coating of crosslinkable olefinic oligomers. Fluorination of the oligomer coated support is effected with gaseous xenon difluoride (XeF 2 ), optionally under inert gas conditions, or with a mixture of fluorine and an inert carrier gas.
- XeF 2 gaseous xenon difluoride
- the composite material thus obtained is suitable for use in the isolation of DNA out of complex mixtures, where apart of DNA also RNA, proteins, low molecular substances and salts are present.
- DNA also RNA, proteins, low molecular substances and salts are present.
- HPLC high pressure
- HIC hydrophobic interaction chromatography
- sorbents on the basis of macroporous silica gels, modified with vinyl monomers like N-vinyl-pyrrolidone, styrene, ethylene, vinylethyl ether (authors certificate, USSR, No. 687081, 1979).
- the sorbent materials are manufactured in applying said monomers to ⁇ -aminopropylated or silanised silica gel and subsequent polymerisation.
- These sorbents possess good mechanical stability, do not swell in solvents and may be used for the separation of proteins and other active biomolecules. No hint is given for using these materials in the separation of nucleic acids.
- the object of the present invention is to provide a sorbent material with improved characteristics in separation and purification capacity and increased selectivity in the specific action of the sorbent material on biopolymers having an advanced surface for biotechnological applications, such as isolation and separation of biopolymers, primarily in aqueous media, with improved access area of the separation surface in a separation medium and improved stability of the coating for the construction of material suitable for chromatographical applications like HPLC and fast sample preparations via solid phase extraction in compact cartridges for PCR-applications.
- the object is solved by a polymer obtainable by a process of polymerizing at least three components [A], [B], and [M] wherein
- [A], [B], and [M] have the same meaning as in claim 1 and x, y, and z are independent of each other an integer of 1-100 and p is a number between 2 and 5000.
- Subject matter of the invention is also a monomer having the structure [A] x —[B] y —[M] z wherein [A], [B], and [M] have the same meaning as in claim 1 and x, y, and z is independent of each other an integer of 1-100
- a chromatographic material comprising essentially a polymer of the invention is also subject matter of the invention as well as a composite sorbent material having a support which is at least partially covered by a polymer coating of the invention.
- a sorbent material having a previously dehydrated solid support having a copolymer coating comprising substituted or unsubstituted aromatic vinyl monomers, substituted or unsubstituted aromatic crosslinking compounds and optionally non-saturated carboxylic acids or esters or non-saturated alcohols or ethers, respectively.
- the coating can be attached to the support via a vinylchlorsilane.
- Substituents of the compounds of the copolymer coating modify the hydrophobic/hydrophilic properties on the surface of the coating and provide an essentially improved selectivity for selected applications.
- the functional groups exhibiting hydrophilic properties provide an essentially better wetting of the inner and outer surfaces of the pores of the sorbent material.
- the support of the sorbent material of the invention is porous inorganic material selected from the group comprising inorganic metal oxides, such as oxides of aluminum, titanium, zirconium, silicon and/or iron.
- inorganic metal oxides such as oxides of aluminum, titanium, zirconium, silicon and/or iron.
- silica gel having an average pore size of 100-2000 ⁇ , preferably 300-1000 ⁇ , and a specific surface of 20-300 m 2 /g, more preferably 20-200 m 2 /g or 20-100 m 2 /g.
- the support containing inorganic materials is in particle-like or monolithic membrane-like form and has a porous structure which shows a bidisperse or oligodisperse distribution of pore sizes.
- Such structures build, e. g., the basis for sorbent materials according to the present invention, which allow additionally to the separation of biopolymers such as nucleic acids the improved retention of low molecular weight substances having, e. g., molecular weights of less than 500 Da, salts and proteins, whereas nucleic acids are passing without retention.
- the separation of nucleic acids, e. g. DNA is conducted in one step.
- Such bidisperse supports may preferentially be obtained by means of gelling (gel building) of silica sols, starting the process with the mixture of two size types of monodisperse colloidal silica particles.
- the mass proportion of these two types of colloidal particles determines the proportion and distribution of differently sized pores in the final silica support material.
- the ratio of the mean diameter of the large pore size distribution and the lower pore size distribution is in the range of 2-15 or 3-15 nm, in particular 4-10 nm.
- the mean diameter of the larger pore size distribution should not be smaller than 25 to 50 nm and should not exceed 200 nm (2000 ⁇ ), preferably 100 nm (1000 ⁇ ).
- Especially preferred mean diameters of the large pore size distribution are in the range of 20-100, more preferably 25-80 nm.
- the mass proportion of the smaller to larger pores represents (90-60, preferably 85-70):(10-40, preferably 15-30) (in %).
- the polymer coating preferably has a thickness of about 10 to 250 Angström, preferably 10 to 100 Angström and micropores of less than 50 ⁇ accessable to water, salts, and low molecular weight substances being non-adsorptive towards nucleic acids and adsorptive towards proteins.
- the previously dried support material is modified by treating it with a boiling solution of a vinylchlorsilane. Treatment of the surface of the support material is conducted at boiling temperature of the solvent used. This treatment enables the following chemosorption of the copolymer and is the basis for enhanced chemical stability and durability of the final sorbent material. Washing of the silanised support, e. g. silica gel, for removal of unreacted silane is done by multiple extraction with organic solvents (ethanol, acetone, toluene, dioxane).
- organic solvents ethanol, acetone, toluene, dioxane
- Polymerisation of the copolymer composition is performed by polymerisation in solution of an organic solvent for a period of 2 to 8 h at the boiling temperature of the solvent or in aqueous solution (emulsion polymerisation) with an emulgator and in the presence of an initiator for radical polymerisation.
- Suitable initiators comprise benzoyl peroxide, potassium peroxodisulfate as a water soluble initiator, ammonium persulfate, etc.
- a 5-20% solution of copolymer is used.
- a mass ratio of silica gel matrix and solution of 1:3-6 is also preferred. Finally unreacted material is removed and the composite material is recovered and dried.
- the sorbent material according to the invention is useful in separation processes, enhancing the ease of handling and the speed of these processes.
- the substances to be separated are nucleic acids and/or proteins.
- a conventionally used chromatographic column or cartridge can be filled, at least partially, with the sorbent material of the invention.
- the sorbent material of the invention behaves similar to other solid chromatographic supports so that the methods for filling chromatographic columns or cartridges can be used in an analogous manner.
- the support for carrying out chromatographic separations can also be provided in the form of a membrane-like item comprising the sorbent material of the invention, wherein the sorbent material is embedded in a polymeric matrix such as a nylon membrane. Also other membrane materials which are used in preparation, isolation or separation of biomolecules can be used as matrix for embedding a sorbent material of the present invention.
- a chromatographic material of the invention it is advantageous to provide the sorbent material according to the invention in bulk or a chromatographic column or cartridge or membrane-like device together with filter materials, reagents and/or buffers or other devices or chemicals for performing fast sample preparations and chromatographic separations.
- This item can especially be provided in form of a kit or a miniaturized device in form of chips or microreactors.
- the chromatographic separation is not limited in its scale. It can be used in any chromatographic operation for separation, isolation, identification, purification and/or detection of biomolecules, in particular nucleic acids, in preparative or analytical scale.
- the present invention provides a product with advanced sorption properties that allows to use this product for chromatography of biopolymers according to the object of the invention.
- the process of the invention leads to a sorbent material with high binding capacity towards proteins and minimal non-specific sorption of nucleic acids, which is accomplished by using copolymers having complementary functional groups.
- the inventive process allows manufacturing of a sorbent with adjusted hydrophilic properties having high mechanical stability and very good hydrodynamic characteristics.
- the sorbent has been obtained similar to example 1, but with a polymer composition without component [M] (2-hydroxyethylmethacrylate).
- the sorbent has been obtained similar to example 3, but with a polymer composition without component [M] (methacrylic acid).
- silanized silica 10 g of macroporous silica gel previously dried in vacuo at 200° C. for 3 h having an average pore diameter of 300 A are treated with a boiling solution of dimethylvinylchlorosilane corresponding to example 1.
- the thus obtained silanized silica will be transferred into a three bottlenecked flask with a stirrer, a thermometer and a reflux condenser.
- To the flask 40 ml toluene, 3 ml chlorovinyl benzene, 0.4 ml divinylbenzene, 0.4 ml allylic alcohol and 0.30 g benzoil peroxide will be added.
- the polymerization is conducted for 6 h at 102° C. The further treatment will be achieved corresponding to example 1.
- the sorbent has been obtained similar to example 5, but with a polymer composition without component [M] (allylic alcohol).
- the ready obtained silica gel was grinded, fractionated and analysed for pore size distribution both by mercury porometry (according to DIN 66 133 (1993)) and BET-method (according to ISO 9277). These analyses showed a preferential pore size in two classes of 5 nm (appr. 85%) and 28 nm (appr. 15%) and a sorption volume of 0.7 cm 3 /g.
- silica gel sorbent was prepared as in example 7, with following variations:
- the porogrammes obtained by testing the sorbents based on the macroporous silica gel show the distribution of the pores in differential and integral manner and allow to determine the medium pore size of the sorbent as well as the effective thickness of the polymeric layer, which is 5-7.5 nm.
- the kit contains all necessary reagents for lysis of cells or tissue and genomic DNA purification.
- the resulting DNA is suitable for most enzymatic reactions (restriction digests, PCR, sequencing etc.).
- DNA flows through the column during a short, one-step purification procedure.
- kits are stable at room temperature during shipment. After arrival store the kit at +2° C. to +8° C. Columns may be stored at room temperature.
- Buffer G1 10 vials blue
- Buffer G2 10 vials blue
- each for 5 isolations Nexttec clean-columns 50 columns
- the kit contains all necessary reagents for lysis of bacterial cells and DNA purification. It is approved for many Gram( ⁇ ) as well as Gram(+) bacteria. The resulting DNA is suitable for most enzymatic reactions (restriction digests, PCR, sequencing etc.).
- DNA flows through the column during a short, one-step purification procedure.
- kit components are stable at room temperature during shipment. After arrival store RNase solution at ⁇ 20° C. The other kit components must be stored at +2° C. to +8° C. Nexttec clean-columns may be stored at room temperature.
- Buffer B1 (basis buffer) 5 vials (white), each for 10 isolations Buffer B2 5 vials (white), each for 10 isolations Buffer B3 5 vials (white), each for 10 isolations Nexttec clean-columns 50 columns RNase solution 1 vial (white), for 50 isolations
- BSA bovine serum albumin
- Salt retention was measured using copper sulphate (CuSO 4 ) as a model salt.
- a solution of this salt in water results in a blue coloured liquid.
- the optical density of the solution is linearly depending on the CuSO 4 concentration.
- the concentration of CuSO 4 in both solutions was then calculated using a linear regression resulting from a standard curve of different CuSO 4 concentrations in water.
- the applied amount of CuSO 4 was set at 100% and the eluted amount of CuSO 4 was recalculated in % of the applied amount. The difference between both values gives the salt retention in %.
- the lysate contains low molecular weight compounds, proteins (peptides), nucleic acids and other compounds.
- the concentration of DNA before (crude lysate) and after (eluate) passing a sorbent containing column was measured using PicoGreen, a fluorescent dye, which binds only to doublestranded DNA, according to the manufacturer's instructions (Molecular Probes c/o Invitrogen).
- PicoGreen a fluorescent dye, which binds only to doublestranded DNA, according to the manufacturer's instructions (Molecular Probes c/o Invitrogen).
- PicoGreen a fluorescent dye, which binds only to doublestranded DNA
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Genetics & Genomics (AREA)
- Inorganic Chemistry (AREA)
- Polymers & Plastics (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Nanotechnology (AREA)
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Crystallography & Structural Chemistry (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Materials Engineering (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
- Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US12/830,676 US20110186519A1 (en) | 2004-04-02 | 2010-07-06 | Process for manufacturing a composite sorbent material for chormatographical separation of biopolymers |
US14/033,074 US20140094596A1 (en) | 2004-04-02 | 2013-09-20 | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers |
US14/966,269 US20160097048A1 (en) | 2004-04-02 | 2015-12-11 | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers |
US15/429,668 US10221411B2 (en) | 2004-04-02 | 2017-02-10 | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers |
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP04008147 | 2004-04-02 | ||
EP04008147.3 | 2004-04-02 | ||
PCT/EP2005/051490 WO2005095476A1 (fr) | 2004-04-02 | 2005-04-01 | Procede de fabrication de materiau sorbant composite pour la separation chromatographique de biopolymeres |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2005/051490 A-371-Of-International WO2005095476A1 (fr) | 2004-04-02 | 2005-04-01 | Procede de fabrication de materiau sorbant composite pour la separation chromatographique de biopolymeres |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US12/830,676 Continuation US20110186519A1 (en) | 2004-04-02 | 2010-07-06 | Process for manufacturing a composite sorbent material for chormatographical separation of biopolymers |
Publications (1)
Publication Number | Publication Date |
---|---|
US20080154029A1 true US20080154029A1 (en) | 2008-06-26 |
Family
ID=34963789
Family Applications (5)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US11/547,011 Abandoned US20080154029A1 (en) | 2004-04-02 | 2005-04-01 | Process For Manufacturing a Composite Sorbent Material For Chromatographical Separation of Biopolymers |
US12/830,676 Abandoned US20110186519A1 (en) | 2004-04-02 | 2010-07-06 | Process for manufacturing a composite sorbent material for chormatographical separation of biopolymers |
US14/033,074 Abandoned US20140094596A1 (en) | 2004-04-02 | 2013-09-20 | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers |
US14/966,269 Abandoned US20160097048A1 (en) | 2004-04-02 | 2015-12-11 | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers |
US15/429,668 Active US10221411B2 (en) | 2004-04-02 | 2017-02-10 | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers |
Family Applications After (4)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US12/830,676 Abandoned US20110186519A1 (en) | 2004-04-02 | 2010-07-06 | Process for manufacturing a composite sorbent material for chormatographical separation of biopolymers |
US14/033,074 Abandoned US20140094596A1 (en) | 2004-04-02 | 2013-09-20 | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers |
US14/966,269 Abandoned US20160097048A1 (en) | 2004-04-02 | 2015-12-11 | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers |
US15/429,668 Active US10221411B2 (en) | 2004-04-02 | 2017-02-10 | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers |
Country Status (5)
Country | Link |
---|---|
US (5) | US20080154029A1 (fr) |
EP (2) | EP2236529B1 (fr) |
JP (1) | JP5173406B2 (fr) |
AT (1) | ATE547442T1 (fr) |
WO (1) | WO2005095476A1 (fr) |
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090301169A1 (en) * | 2008-05-02 | 2009-12-10 | Naval Research Laboratory | Selective membranes/thin films for analytical applications |
WO2016040697A1 (fr) | 2014-09-10 | 2016-03-17 | Quantumdx Group Limited | Matériau sorbant pour la séparation de bio-macromolécules |
CN109627402A (zh) * | 2018-12-14 | 2019-04-16 | 江南大学 | 一种聚苯乙烯基高折射率纳米复合材料的制备方法 |
US10695744B2 (en) | 2015-06-05 | 2020-06-30 | W. R. Grace & Co.-Conn. | Adsorbent biprocessing clarification agents and methods of making and using the same |
US11229896B2 (en) | 2014-01-16 | 2022-01-25 | W.R. Grace & Co.—Conn. | Affinity chromatography media and chromatography devices |
US11376561B2 (en) * | 2018-10-02 | 2022-07-05 | Waters Technologies Corporation | Sorbent particles for sample treatment |
US11389783B2 (en) | 2014-05-02 | 2022-07-19 | W.R. Grace & Co.-Conn. | Functionalized support material and methods of making and using functionalized support material |
US11529610B2 (en) | 2012-09-17 | 2022-12-20 | W.R. Grace & Co.-Conn. | Functionalized particulate support material and methods of making and using the same |
US11628381B2 (en) | 2012-09-17 | 2023-04-18 | W.R. Grace & Co. Conn. | Chromatography media and devices |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP1620479B1 (fr) | 2002-10-15 | 2013-07-24 | ExxonMobil Chemical Patents Inc. | Compositions à base d'adhesifs polyolefiniques et articles fabriques à partir de celles-ci |
JP5097506B2 (ja) | 2007-11-05 | 2012-12-12 | 日東電工株式会社 | 水酸基を有する多孔質樹脂粒子の製造方法 |
CN104174425B (zh) * | 2014-07-11 | 2017-08-11 | 中国科学院生态环境研究中心 | 一种用于挥发性有机物催化氧化的催化剂及其制备方法 |
GB201703625D0 (en) | 2017-03-07 | 2017-04-19 | Quantumdx Group Ltd | Aptamer based methods for protein detection |
CN107626278A (zh) * | 2017-09-04 | 2018-01-26 | 无锡普爱德环保科技有限公司 | 新型硅胶吸附材料的制备方法 |
EP4021639A1 (fr) | 2019-08-30 | 2022-07-06 | Boehringer Ingelheim Vetmedica GmbH | Instrument de filtration, kit et procédé destinés au prétraitement d'un échantillon |
CL2020002629A1 (es) | 2020-10-12 | 2021-01-15 | Taag Genetics S A | Nuevo dispositivo de biología molecular para extracción y purificación de ácidos nucleicos desde diferentes tipos de muestras biológicas que comprende resinas de adsorción |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4045353A (en) * | 1974-10-15 | 1977-08-30 | Toyo Soda Manufacturing Co., Ltd. | High-energy radiation induced polymerization on a chromatographic solid support |
US4600646A (en) * | 1984-08-15 | 1986-07-15 | E. I. Du Pont De Nemours And Company | Metal oxide stabilized chromatography packings |
US4732887A (en) * | 1984-10-12 | 1988-03-22 | Asahi Kasei Kogyo Kabushiki Kaisha | Composite porous material, process for production and separation of metallic element |
US5163994A (en) * | 1987-03-11 | 1992-11-17 | Basf Aktiengesellschaft | Crop protection agent containing an active ingredient |
US5750258A (en) * | 1994-04-11 | 1998-05-12 | Ube Nitto Kasei Co., Ltd. | Crosslinked resin-coated silica fine particles and process for the production thereof |
US6316527B1 (en) * | 1999-09-16 | 2001-11-13 | Rohm And Haas Company | Modified SAN resin blend compositions and articles produced therefrom |
US6417239B1 (en) * | 1997-08-28 | 2002-07-09 | Eastman Chemical Company | Methods of making modified condensation polymers |
US20030134938A1 (en) * | 2001-09-19 | 2003-07-17 | Dainichiseika Color & Chem. Mfg Co., Ltd. | Process for producing sulfonated solid particles |
Family Cites Families (23)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5923564B2 (ja) | 1980-03-03 | 1984-06-02 | 昭光通商株式会社 | 三次元架橋高分子を主体とする多孔質体の成形用組成物 |
GB2191110B (en) * | 1986-06-06 | 1989-12-06 | Plessey Co Plc | Chromatographic separation device |
DE3707692A1 (de) * | 1987-03-11 | 1988-09-22 | Basf Ag | Polymere traeger fuer pestizide |
JP2790381B2 (ja) * | 1990-02-03 | 1998-08-27 | 三井化学株式会社 | 有芯多層構造エマルション粒子 |
JPH0596184A (ja) * | 1991-10-02 | 1993-04-20 | Showa Denko Kk | 弱酸性陽イオン交換体 |
JP3217417B2 (ja) * | 1991-12-24 | 2001-10-09 | 三井化学株式会社 | 中空エマルション粒子 |
US6420127B1 (en) | 1994-11-18 | 2002-07-16 | Washington University | Compounds and pharmaceutical compositions for the treatment and prophylaxis of bacterial infections |
DK0880536T4 (en) * | 1996-02-06 | 2015-09-07 | Roche Diagnostics Gmbh | A process for the preparation of purified nucleic acid and the use thereof |
US5792331A (en) * | 1996-12-19 | 1998-08-11 | Dionex Corporation | Preformed polymer coating process and product |
US6056877A (en) * | 1997-12-05 | 2000-05-02 | Transgenomic, Inc. | Non-polar media for polynucleotide separations |
US5986020A (en) * | 1997-08-05 | 1999-11-16 | Campbell; J. David | Process for producing hyperbranched polymers |
BR9811372A (pt) * | 1997-08-28 | 2000-08-29 | Eastman Chem Co | Composições de látex diol, de revestimento, e de veìculo de tinta, e, processo para a preparação da comoposição de látex diol |
JP4037537B2 (ja) * | 1998-09-01 | 2008-01-23 | 積水化学工業株式会社 | 液体クロマトグラフィー用充填剤 |
JP4074393B2 (ja) * | 1998-11-09 | 2008-04-09 | 積水化学工業株式会社 | 液体クロマトグラフィー用充填剤 |
EP1020220A1 (fr) | 1999-01-11 | 2000-07-19 | Mira Diagnostica GmbH | Nouveau polymere contenant des groupements de fluor |
US6824866B1 (en) * | 1999-04-08 | 2004-11-30 | Affymetrix, Inc. | Porous silica substrates for polymer synthesis and assays |
WO2001074482A1 (fr) * | 2000-04-05 | 2001-10-11 | Japan Chemical Innovation Institute | Nouveau materiau utilise pour la separation et procede de separation |
US6548264B1 (en) * | 2000-05-17 | 2003-04-15 | University Of Florida | Coated nanoparticles |
JP2002194025A (ja) | 2000-12-22 | 2002-07-10 | Sumitomo Chem Co Ltd | エチルスチレン・ジビニルベンゼン共重合体および該重合体からなるアミノ酸用分離材。 |
JP3957179B2 (ja) * | 2001-09-18 | 2007-08-15 | オルガノ株式会社 | 有機多孔質イオン交換体 |
JP2003165926A (ja) * | 2001-09-19 | 2003-06-10 | Dainichiseika Color & Chem Mfg Co Ltd | スルホン化固体粒子の製造方法 |
US6991852B2 (en) * | 2002-03-13 | 2006-01-31 | Regents Of The University Of Minnesota | Silica-based materials and methods |
US20060243658A1 (en) | 2002-11-08 | 2006-11-02 | Zubov Vitali P | Sorbent material having a covalently attached perfluorinated surface with functional groups |
-
2005
- 2005-04-01 US US11/547,011 patent/US20080154029A1/en not_active Abandoned
- 2005-04-01 WO PCT/EP2005/051490 patent/WO2005095476A1/fr active Application Filing
- 2005-04-01 JP JP2007505564A patent/JP5173406B2/ja active Active
- 2005-04-01 EP EP10163239A patent/EP2236529B1/fr active Active
- 2005-04-01 AT AT05731668T patent/ATE547442T1/de active
- 2005-04-01 EP EP05731668A patent/EP1756178B1/fr active Active
-
2010
- 2010-07-06 US US12/830,676 patent/US20110186519A1/en not_active Abandoned
-
2013
- 2013-09-20 US US14/033,074 patent/US20140094596A1/en not_active Abandoned
-
2015
- 2015-12-11 US US14/966,269 patent/US20160097048A1/en not_active Abandoned
-
2017
- 2017-02-10 US US15/429,668 patent/US10221411B2/en active Active
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4045353A (en) * | 1974-10-15 | 1977-08-30 | Toyo Soda Manufacturing Co., Ltd. | High-energy radiation induced polymerization on a chromatographic solid support |
US4600646A (en) * | 1984-08-15 | 1986-07-15 | E. I. Du Pont De Nemours And Company | Metal oxide stabilized chromatography packings |
US4732887A (en) * | 1984-10-12 | 1988-03-22 | Asahi Kasei Kogyo Kabushiki Kaisha | Composite porous material, process for production and separation of metallic element |
US5163994A (en) * | 1987-03-11 | 1992-11-17 | Basf Aktiengesellschaft | Crop protection agent containing an active ingredient |
US5750258A (en) * | 1994-04-11 | 1998-05-12 | Ube Nitto Kasei Co., Ltd. | Crosslinked resin-coated silica fine particles and process for the production thereof |
US6417239B1 (en) * | 1997-08-28 | 2002-07-09 | Eastman Chemical Company | Methods of making modified condensation polymers |
US6316527B1 (en) * | 1999-09-16 | 2001-11-13 | Rohm And Haas Company | Modified SAN resin blend compositions and articles produced therefrom |
US20030134938A1 (en) * | 2001-09-19 | 2003-07-17 | Dainichiseika Color & Chem. Mfg Co., Ltd. | Process for producing sulfonated solid particles |
Cited By (15)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090301169A1 (en) * | 2008-05-02 | 2009-12-10 | Naval Research Laboratory | Selective membranes/thin films for analytical applications |
US8148161B2 (en) | 2008-05-02 | 2012-04-03 | The United States Of America, As Represented By The Secretary Of The Navy | Selective membranes/thin films for analytical applications |
US11628381B2 (en) | 2012-09-17 | 2023-04-18 | W.R. Grace & Co. Conn. | Chromatography media and devices |
US11529610B2 (en) | 2012-09-17 | 2022-12-20 | W.R. Grace & Co.-Conn. | Functionalized particulate support material and methods of making and using the same |
US11229896B2 (en) | 2014-01-16 | 2022-01-25 | W.R. Grace & Co.—Conn. | Affinity chromatography media and chromatography devices |
US11389783B2 (en) | 2014-05-02 | 2022-07-19 | W.R. Grace & Co.-Conn. | Functionalized support material and methods of making and using functionalized support material |
US10792640B2 (en) | 2014-09-10 | 2020-10-06 | Quantumdx Group Limited | Sorbent material for separating bio-macromolecules |
AU2015314928B2 (en) * | 2014-09-10 | 2020-07-02 | Quantumdx Group Limited | Sorbent material for separating bio-macromolecules |
EP3191590A4 (fr) * | 2014-09-10 | 2018-08-01 | Quantumdx Group Limited | Matériau sorbant pour la séparation de bio-macromolécules |
US20170282154A1 (en) * | 2014-09-10 | 2017-10-05 | Quantumdx Group Limited | Sorbent material for separating bio-macromolecules |
CN106604992A (zh) * | 2014-09-10 | 2017-04-26 | 康特姆斯集团有限公司 | 用于分离生物大分子的吸附剂材料 |
WO2016040697A1 (fr) | 2014-09-10 | 2016-03-17 | Quantumdx Group Limited | Matériau sorbant pour la séparation de bio-macromolécules |
US10695744B2 (en) | 2015-06-05 | 2020-06-30 | W. R. Grace & Co.-Conn. | Adsorbent biprocessing clarification agents and methods of making and using the same |
US11376561B2 (en) * | 2018-10-02 | 2022-07-05 | Waters Technologies Corporation | Sorbent particles for sample treatment |
CN109627402A (zh) * | 2018-12-14 | 2019-04-16 | 江南大学 | 一种聚苯乙烯基高折射率纳米复合材料的制备方法 |
Also Published As
Publication number | Publication date |
---|---|
US20110186519A1 (en) | 2011-08-04 |
US10221411B2 (en) | 2019-03-05 |
JP2007530760A (ja) | 2007-11-01 |
EP1756178B1 (fr) | 2012-02-29 |
EP2236529B1 (fr) | 2012-05-23 |
EP2236529A1 (fr) | 2010-10-06 |
ATE547442T1 (de) | 2012-03-15 |
US20140094596A1 (en) | 2014-04-03 |
JP5173406B2 (ja) | 2013-04-03 |
WO2005095476A1 (fr) | 2005-10-13 |
US20160097048A1 (en) | 2016-04-07 |
EP1756178A1 (fr) | 2007-02-28 |
US20170152501A1 (en) | 2017-06-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US10221411B2 (en) | Process for manufacturing a composite sorbent material for chromatographical separation of biopolymers | |
JP3395906B2 (ja) | 核酸断片の分離法 | |
US9624252B2 (en) | Selective nucleic acid fragment recovery | |
US8569477B2 (en) | Method for isolating nucleic acids comprising the use of ethylene glycol multimers | |
US8288169B2 (en) | Surface mediated self-assembly of nanoparticles | |
JP2839144B2 (ja) | 複合ポリマー、それらの製造、及び液相クロマトグラフイーにおけるそれらの使用 | |
US20090048439A1 (en) | Isolation of nucleic acids molecules using modified solid supports | |
JP2011503244A (ja) | 核酸の単離方法またはリン酸化タンパク質の単離方法における粒子およびその使用 | |
US20140287416A1 (en) | Polyelectrolyte-coated size-exclusion ion-exchange particles | |
JP2001204462A (ja) | 核酸の抽出方法 | |
JP2022515769A (ja) | 大孔径アガロース | |
JP2006512457A (ja) | 収着剤、及びプラスミドdnaの分離方法 | |
WO2005079984A1 (fr) | Particules echangeuse d'ions d'exclusion enduites de polyelectrolytes | |
US20050196856A1 (en) | Polyelectrolyte-coated size-exclusion ion-exchange particles | |
JP4929635B2 (ja) | マレイミド基含有多孔質架橋ポリスチレン粒子及びその製造方法 | |
Li et al. | High-speed chromatographic purification of plasmid DNA with a customized biporous hydrophobic adsorbent | |
US20060243658A1 (en) | Sorbent material having a covalently attached perfluorinated surface with functional groups | |
US7772152B2 (en) | Composite polymer-coated sorbent with a bidisperse pore size distribution for the simultaneous separation and desalting of biopolymers | |
EP1848793B1 (fr) | Particules d'echange ionique revetues de polyelectrolytes | |
US20070207460A1 (en) | Method For Enriching And Stabilising Components Which Contain Dna And Which Are Made Of Biological Materials | |
US20040002594A1 (en) | Removal of extraneous substances from biological fluids containing nucleic acids and the recovery of nucleic acids |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
AS | Assignment |
Owner name: NEXTEC GMBH, GERMANY Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:BALAYAN, HAMLET;LEISER, ROBERT-MATTHIAS;PLOBNER, LUTZ;AND OTHERS;REEL/FRAME:018727/0809;SIGNING DATES FROM 20060922 TO 20060926 |
|
AS | Assignment |
Owner name: NEXTTEC GMBH, GERMANY Free format text: CORRECTIVE ASSIGNMENT TO CORRECT THE RECEIVING PARTY, PREVIOUSLY RECORDED AT REEL 018727, FRAME 0809.;ASSIGNORS:BALAYAN, HAMLET;LEISER, ROBERT-MATTHIAS;PLOBNER, LUTZ;AND OTHERS;REEL/FRAME:020863/0500;SIGNING DATES FROM 20060922 TO 20060926 |
|
STCB | Information on status: application discontinuation |
Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION |