CA2474436C - Process for the coarse clarification of lysed cell material from microorganisms - Google Patents
Process for the coarse clarification of lysed cell material from microorganisms Download PDFInfo
- Publication number
- CA2474436C CA2474436C CA2474436A CA2474436A CA2474436C CA 2474436 C CA2474436 C CA 2474436C CA 2474436 A CA2474436 A CA 2474436A CA 2474436 A CA2474436 A CA 2474436A CA 2474436 C CA2474436 C CA 2474436C
- Authority
- CA
- Canada
- Prior art keywords
- lysate
- pressure
- normal atmospheric
- atmospheric pressure
- phase
- 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.)
- Expired - Fee Related
Links
- 238000000034 method Methods 0.000 title claims abstract description 31
- 244000005700 microbiome Species 0.000 title claims abstract description 12
- 238000005352 clarification Methods 0.000 title abstract description 6
- 239000000463 material Substances 0.000 title abstract description 4
- 239000006166 lysate Substances 0.000 claims abstract description 20
- 239000012071 phase Substances 0.000 claims abstract description 17
- 239000002244 precipitate Substances 0.000 claims abstract description 12
- 239000013592 cell lysate Substances 0.000 claims abstract description 7
- 239000007791 liquid phase Substances 0.000 claims abstract description 6
- 230000002934 lysing effect Effects 0.000 claims abstract description 3
- 241000894006 Bacteria Species 0.000 claims description 6
- 102000039446 nucleic acids Human genes 0.000 abstract description 3
- 108020004707 nucleic acids Proteins 0.000 abstract description 3
- 150000007523 nucleic acids Chemical class 0.000 abstract description 3
- 239000000872 buffer Substances 0.000 description 22
- 239000013612 plasmid Substances 0.000 description 15
- 230000009089 cytolysis Effects 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 8
- 238000000926 separation method Methods 0.000 description 7
- 238000001914 filtration Methods 0.000 description 6
- 238000006386 neutralization reaction Methods 0.000 description 6
- 239000002028 Biomass Substances 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 239000000470 constituent Substances 0.000 description 5
- 239000002158 endotoxin Substances 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 239000011541 reaction mixture Substances 0.000 description 5
- 210000002421 cell wall Anatomy 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 3
- 239000008004 cell lysis buffer Substances 0.000 description 3
- 238000005191 phase separation Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 239000008346 aqueous phase Substances 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000012139 lysis buffer Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- SCVFZCLFOSHCOH-UHFFFAOYSA-M potassium acetate Chemical compound [K+].CC([O-])=O SCVFZCLFOSHCOH-UHFFFAOYSA-M 0.000 description 2
- ONQDVAFWWYYXHM-UHFFFAOYSA-M potassium lauryl sulfate Chemical compound [K+].CCCCCCCCCCCCOS([O-])(=O)=O ONQDVAFWWYYXHM-UHFFFAOYSA-M 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000004062 sedimentation Methods 0.000 description 2
- 238000011179 visual inspection Methods 0.000 description 2
- WZFUQSJFWNHZHM-UHFFFAOYSA-N 2-[4-[2-(2,3-dihydro-1H-inden-2-ylamino)pyrimidin-5-yl]piperazin-1-yl]-1-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethanone Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)N1CCN(CC1)CC(=O)N1CC2=C(CC1)NN=N2 WZFUQSJFWNHZHM-UHFFFAOYSA-N 0.000 description 1
- 101000743092 Bacillus spizizenii (strain DSM 15029 / JCM 12233 / NBRC 101239 / NRRL B-23049 / TU-B-10) tRNA3(Ser)-specific nuclease WapA Proteins 0.000 description 1
- 101000743093 Bacillus subtilis subsp. natto (strain BEST195) tRNA(Glu)-specific nuclease WapA Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 238000011021 bench scale process Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 230000003311 flocculating effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 235000011056 potassium acetate Nutrition 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
- 238000005406 washing 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
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Crystallography & Structural Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Saccharide Compounds (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE10207170.5 | 2002-02-20 | ||
| DE10207170A DE10207170A1 (de) | 2002-02-20 | 2002-02-20 | Verfahren zur Grobklärung von Zellaufschlüssen aus Mikroorganismen |
| PCT/EP2003/001769 WO2003070942A1 (de) | 2002-02-20 | 2003-02-20 | Verfahren zur grobklärung von zellaufschlüssen aus mikroorganismen |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2474436A1 CA2474436A1 (en) | 2003-08-28 |
| CA2474436C true CA2474436C (en) | 2011-08-23 |
Family
ID=27674782
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2474436A Expired - Fee Related CA2474436C (en) | 2002-02-20 | 2003-02-20 | Process for the coarse clarification of lysed cell material from microorganisms |
Country Status (7)
| Country | Link |
|---|---|
| US (1) | US7214508B2 (enExample) |
| EP (1) | EP1478749B1 (enExample) |
| JP (1) | JP4344245B2 (enExample) |
| AT (1) | ATE464380T1 (enExample) |
| CA (1) | CA2474436C (enExample) |
| DE (2) | DE10207170A1 (enExample) |
| WO (1) | WO2003070942A1 (enExample) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2498518A1 (en) | 2002-09-13 | 2004-03-25 | Valentis, Inc. | Apparatus and method for preparative scale purification of nucleic acids |
| EP2088196A1 (en) * | 2008-02-08 | 2009-08-12 | Boehringer Ingelheim RCV GmbH & Co KG | Methods and devices for producing biomolecules |
| US11235323B2 (en) | 2008-08-27 | 2022-02-01 | Life Technologies Corporation | Apparatus for and method of processing biological samples |
| JP5492207B2 (ja) | 2008-08-27 | 2014-05-14 | ライフ テクノロジーズ コーポレーション | 生物学的サンプルの処理装置および処理方法 |
| GB2473868A (en) | 2009-09-28 | 2011-03-30 | Invitrogen Dynal As | Apparatus and method of automated processing of biological samples |
| US9724460B2 (en) | 2014-03-25 | 2017-08-08 | Oakwood Healthcare, Inc. | Controlled nucleation from gas-supersaturated liquid |
| WO2021126818A1 (en) | 2019-12-18 | 2021-06-24 | Life Technologies Corporation | Systems, methods, and devices for automated nucleic acid and protein isolation |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4143639C2 (de) * | 1991-12-02 | 2002-10-24 | Qiagen Gmbh | Verfahren zur Isolierung und Reinigung von Nukleinsäuren |
| JP2002507121A (ja) * | 1997-06-25 | 2002-03-05 | プロメガ コーポレイション | Ｒｎａの単離方法 |
| US6120985A (en) * | 1997-10-31 | 2000-09-19 | Bbi Bioseq, Inc. | Pressure-enhanced extraction and purification |
| CA2498518A1 (en) * | 2002-09-13 | 2004-03-25 | Valentis, Inc. | Apparatus and method for preparative scale purification of nucleic acids |
-
2002
- 2002-02-20 DE DE10207170A patent/DE10207170A1/de not_active Withdrawn
-
2003
- 2003-02-20 US US10/504,258 patent/US7214508B2/en not_active Expired - Lifetime
- 2003-02-20 EP EP03706546A patent/EP1478749B1/de not_active Expired - Lifetime
- 2003-02-20 AT AT03706546T patent/ATE464380T1/de active
- 2003-02-20 CA CA2474436A patent/CA2474436C/en not_active Expired - Fee Related
- 2003-02-20 JP JP2003569835A patent/JP4344245B2/ja not_active Expired - Fee Related
- 2003-02-20 WO PCT/EP2003/001769 patent/WO2003070942A1/de not_active Ceased
- 2003-02-20 DE DE50312615T patent/DE50312615D1/de not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| US7214508B2 (en) | 2007-05-08 |
| DE10207170A1 (de) | 2003-09-04 |
| EP1478749B1 (de) | 2010-04-14 |
| CA2474436A1 (en) | 2003-08-28 |
| DE50312615D1 (de) | 2010-05-27 |
| WO2003070942A1 (de) | 2003-08-28 |
| JP2005517444A (ja) | 2005-06-16 |
| JP4344245B2 (ja) | 2009-10-14 |
| EP1478749A1 (de) | 2004-11-24 |
| ATE464380T1 (de) | 2010-04-15 |
| AU2003208740A1 (en) | 2003-09-09 |
| US20050089950A1 (en) | 2005-04-28 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| MKLA | Lapsed |
Effective date: 20200220 |