DK2631297T3 - Fremgangsmåde, vektorer og manipulerede cellelinjer til at forbedre transfektion i stor målestok - Google Patents
Fremgangsmåde, vektorer og manipulerede cellelinjer til at forbedre transfektion i stor målestok Download PDFInfo
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- DK2631297T3 DK2631297T3 DK12192258.7T DK12192258T DK2631297T3 DK 2631297 T3 DK2631297 T3 DK 2631297T3 DK 12192258 T DK12192258 T DK 12192258T DK 2631297 T3 DK2631297 T3 DK 2631297T3
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- Prior art keywords
- gly
- pro
- arg
- ala
- cells
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
- C07K2319/71—Fusion polypeptide containing domain for protein-protein interaction containing domain for transcriptional activaation, e.g. VP16
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- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/16011—Herpesviridae
- C12N2710/16211—Lymphocryptovirus, e.g. human herpesvirus 4, Epstein-Barr Virus
- C12N2710/16222—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
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- C12N2710/00011—Details
- C12N2710/16011—Herpesviridae
- C12N2710/16611—Simplexvirus, e.g. human herpesvirus 1, 2
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- C12N2800/00—Nucleic acids vectors
- C12N2800/10—Plasmid DNA
- C12N2800/108—Plasmid DNA episomal vectors
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- C12N2800/00—Nucleic acids vectors
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Claims (9)
1. Ekspressionssystem til stabil ekspression af et gen af interesse, hvilket ekspressionssystem omfatter: en første vektor med en Epstein-Barr-virus nuklear antigen-1-(EBNA1) nukleotidsekvens, der koder for et protein, der består af EBNAlc, EBNAlt eller EBNAls som vist i Figur 1, en promoter og et polyadenyleringssignal for EBNAl-nukleotidsekvensen og en oriP-nukleotidsekvens; og en anden vektor med et gen af interesse, en promoter og et polyadenyleringssignal for genet af interesse og en oriP-nukleotidsekvens.
2. Ekspressionssystem ifølge krav 1, hvor EBNA1-proteinet består af SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 11 eller SEQ ID NO: 12.
3. Ekspressionssystem ifølge krav 1 eller 2, hvor EBNAl-nukleotidsekvensen er kodon-optimeret.
4. Ekspressionssystem ifølge krav 1, hvor Epstcin-Barr-virus nuklear antigcn-1- (EBNA1) nukleotidsekvensen består af SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 13 eller SEQ ID NO: 14.
5. Ekspressionssystem ifølge et hvilket som helst af de foregående krav, hvor oriP-nukleotidsekvenseme i den første og den anden vektor uafhængigt omfatter SEQ ID NO: 9 eller SEQ ID NO: 10.
6. Fremgangsmåde in vitro til stabil ekspression af et gen af interesse i pattedyreceller, hvilken fremgangsmåde omfatter: transfektion af en pattedyrecelle med et ekspressionssystem som defineret i et hvilket som helst af de foregående krav; og replikation af cellen for at tilvejebringe pattedyreceller, der stabilt udtrykker genet af interesse.
7. Fremgangsmåde ifølge krav 6, hvor transfektion af cellen med den første og den anden vektor udføres samtidigt, eller cellen transficeres med én af vektorerne først for at frembringe en stabil klon efterfulgt af transfektion med den anden vektor for at frembringe en klon, der stabilt udtrykker genet af interesse.
8. Fremgangsmåde ifølge krav 6 eller 7, hvor pattedyrecelleme er udvalgt fra gruppen bestående af kinesiske hamsterovarie- (CHO) celler, humane embryonyreceller 293 (HEK293) og Madin-Darby-hundenyre- (MDCK) celler.
9. Fremgangsmåde ifølge et hvilket som helst af kravene 6 til 8, hvor pattedyrecelleme er kinesiske hamsterovarie- (CHO) celler.
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| EP09745326A EP2285969B1 (en) | 2008-05-15 | 2009-03-09 | Use of protein kinase b and valproic acid to increase heterologous gene expression in mammalian cells |
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| DK09745326.0T DK2285969T3 (da) | 2008-05-15 | 2009-03-09 | Anvendelse af proteinkinase b og valproinsyre for at forøge heterolog genekspression i pattedyrsceller |
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| US8822214B2 (en) | 2009-09-15 | 2014-09-02 | Medimmune Limited | Cells for transient expression and uses thereof |
| WO2014012082A2 (en) | 2012-07-13 | 2014-01-16 | Zymeworks Inc. | Multivalent heteromultimer scaffold design an constructs |
| KR102332303B1 (ko) | 2013-05-31 | 2021-11-26 | 자임워크스 인코포레이티드 | 감소되거나 침묵화된 효과기 기능을 갖는 이종다량체 |
| AU2014331476B2 (en) * | 2013-10-04 | 2020-07-02 | Advance Sentry Corporation | Methods and devices for nasopharyngeal carcinoma screening |
| CN104707135B (zh) * | 2013-12-11 | 2017-11-07 | 深圳先进技术研究院 | 重组蛋白疫苗、含编码该重组蛋白疫苗的基因的重组表达载体及其应用 |
| SG11201907321TA (en) | 2017-02-07 | 2019-09-27 | Daiichi Sankyo Co Ltd | Anti-gprc5d antibody and molecule comprising the antibody |
| CN106987559A (zh) * | 2017-03-22 | 2017-07-28 | 上海药明生物技术有限公司 | 一种重组chok1细胞株的构建方法及其应用 |
| US20220169997A1 (en) | 2019-04-12 | 2022-06-02 | Therapten Biosciences Inc. | Compositions and methods to manufacture tumor suppressor fusions |
| JP7692905B2 (ja) * | 2019-10-30 | 2025-06-16 | ナショナル リサーチ カウンシル オブ カナダ | 哺乳動物細胞からのタンパク質の生産性を増加させるためのウイルス複製起点 |
| CN113025648B (zh) * | 2019-12-09 | 2023-06-02 | 广东菲鹏制药股份有限公司 | Expi293细胞瞬时表达目标蛋白的方法及其应用 |
| EP4284828A1 (en) | 2021-01-28 | 2023-12-06 | Talem Therapeutics LLC | Anti-sars-cov-2 spike glycoprotein antibodies and the therapeutic use thereof |
| CN119654335A (zh) * | 2022-07-29 | 2025-03-18 | 上海药明生物技术有限公司 | 用于哺乳动物细胞中蛋白质表达的功能增强的工程改造的ebna1 |
| WO2024081918A1 (en) | 2022-10-14 | 2024-04-18 | Talem Therapeutics Llc | Anti-trkb/cd3 antibodies and uses thereof |
| WO2026036019A1 (en) * | 2024-08-09 | 2026-02-12 | Life Technologies Corporation | Method of producing a recombinant protein in hek-293 cells |
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| FR2722506B1 (fr) | 1994-07-13 | 1996-08-14 | Rhone Poulenc Rorer Sa | Composition contenant des acides nucleiques, preparation et utilisations |
| US6797494B1 (en) | 1996-08-16 | 2004-09-28 | Medical Research Council | Self-replicating episomal expression vectors conferring tissue-specific gene expression |
| JP2002505586A (ja) | 1997-06-19 | 2002-02-19 | ザ ジェネラル ホスピタル コーポレーション | 小型エプスタイン・バーウイルスレプリコン |
| US6960429B2 (en) * | 1997-11-12 | 2005-11-01 | Wisconsin Alumni Research Foundation | Inhibition of viral gene activities |
| US5976807A (en) | 1998-03-18 | 1999-11-02 | Pharmacopeia, Inc. | Eukaryotic cells stably expressing genes from multiple transfected episomes |
| US6417002B1 (en) * | 1999-02-11 | 2002-07-09 | Pharmacopeia, Inc. | Method for maintenance and selection of episomes |
| US7723102B2 (en) | 2000-09-28 | 2010-05-25 | Bayer Corporation | Enhanced transfection system |
| AU2002257440A1 (en) | 2001-05-07 | 2002-11-18 | National Research Council Of Canada | Enhanced production of recombinant proteins by transient transfection of suspension-growing mammalian cells |
| US7919270B2 (en) | 2003-09-09 | 2011-04-05 | Acyte Biotech Pty Ltd | Mammalian expression system |
| US7465580B2 (en) | 2004-05-19 | 2008-12-16 | Wisconsin Alumni Research Foundation | Non-cytotoxic oriP replicon |
| EP1861498A4 (en) | 2005-03-17 | 2009-06-24 | Ca Nat Research Council | EXPRESSION VECTORS FOR TRANSIENT GENETIC EXPRESSION AND MAMMALIAN CELLS EXPRESSING THE SAME |
| WO2007048601A1 (en) | 2005-10-28 | 2007-05-03 | F. Hoffmann-La Roche Ag | Protein expression in rodent cells |
| FR2898487B1 (fr) | 2006-03-14 | 2008-11-14 | Spineart Sa Sa | Protheses de disques intervertebrales |
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Also Published As
| Publication number | Publication date |
|---|---|
| US9353382B2 (en) | 2016-05-31 |
| EP2631297A1 (en) | 2013-08-28 |
| US20140051170A1 (en) | 2014-02-20 |
| EP2631297B1 (en) | 2017-04-12 |
| CA2722835C (en) | 2018-11-27 |
| CA2722835A1 (en) | 2009-11-19 |
| EP2285969A4 (en) | 2011-11-16 |
| WO2009137911A1 (en) | 2009-11-19 |
| US8637315B2 (en) | 2014-01-28 |
| DK2285969T3 (da) | 2013-04-08 |
| CA3020928A1 (en) | 2009-11-19 |
| EP2285969A1 (en) | 2011-02-23 |
| US20110039339A1 (en) | 2011-02-17 |
| EP2285969B1 (en) | 2013-01-16 |
| CA3020928C (en) | 2023-08-08 |
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