RU2018128968A - Продуцирование aav в клетках насекомых, способы и композиции для этого - Google Patents
Продуцирование aav в клетках насекомых, способы и композиции для этого Download PDFInfo
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- RU2018128968A RU2018128968A RU2018128968A RU2018128968A RU2018128968A RU 2018128968 A RU2018128968 A RU 2018128968A RU 2018128968 A RU2018128968 A RU 2018128968A RU 2018128968 A RU2018128968 A RU 2018128968A RU 2018128968 A RU2018128968 A RU 2018128968A
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- expression cassette
- baculovirus
- aav
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- rep
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- 241000238631 Hexapoda Species 0.000 title claims 11
- 238000000034 method Methods 0.000 title claims 9
- 238000004519 manufacturing process Methods 0.000 title claims 2
- 239000000203 mixture Substances 0.000 title 1
- 241000701447 unidentified baculovirus Species 0.000 claims 21
- 238000000338 in vitro Methods 0.000 claims 4
- 238000004113 cell culture Methods 0.000 claims 3
- 238000011534 incubation Methods 0.000 claims 3
- 108091081024 Start codon Proteins 0.000 claims 2
- 108700019146 Transgenes Proteins 0.000 claims 2
- 208000015181 infectious disease Diseases 0.000 claims 1
- 238000001890 transfection Methods 0.000 claims 1
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Claims (32)
1. Бакуловирусный вектор, содержащий:
кассету экспрессии AAV Сар;
кассету экспрессии AAV Rep; и
гомологичную область (hr) бакуловируса, расположенную примерно на расстоянии до около 4 тыс. п.о. от стартового кодона кассеты экспрессии AAV.
2. Вектор по п. 1, содержащий в направлении 5'-3' кассету экспрессии Сар, кассету экспрессии Rep и гомологичную область (hr) бакуловируса.
3. Вектор по п. 1, содержащий в направлении 5'-3' кассету экспрессии Rep, кассету экспрессии Сар и гомологичную область (hr) бакуловируса.
4. Вектор по п. 1, содержащий в направлении 5'-3' кассету экспрессии Сар, гомологичную область (hr) бакуловируса и кассету экспрессии Rep.
5. Вектор по п. 1, содержащий в направлении 5'-3' кассету экспрессии Rep, гомологичную область (hr) бакуловируса и кассету экспрессии Сар.
6. Вектор по п. 1, содержащий в направлении 5'-3' гомологичную область (hr) бакуловируса, кассету экспрессии Сар и кассету экспрессии Rep.
7. Вектор по п. 1, содержащий в направлении 5'-3' гомологичную область (hr) бакуловируса, кассету экспрессии Rep и кассету экспрессии Сар.
8. Вектор по п. 1, в котором область hr находится между кассетой экспрессии Rep и кассетой экспрессии Сар, и в котором кассета экспрессии Rep и кассета экспрессии Сар находятся в ориентации голова-к-голове (5'-5').
9. Вектор по любому из пп. 1-8, отличающийся тем, что гомологичная область бакуловируса представляет собой последовательность hr2.
10. Вектор по любому из пп. 1-9, отличающийся тем, что вектор не содержит Rep-связывающий элемент (RBE).
11. Линия клеток насекомых, содержащая клетки, содержащие вектор по любому из пп. 1-10.
12. Линия клеток насекомых по п. 11, отличающаяся тем, что указанные клетки дополнительно содержат второй вектор, причем указанный второй вектор содержит трансген, фланкированный ITR AAV.
13. Способ выращивания бакуловируса in vitro, включающий:
обеспечение культуры клеток насекомых по п. 11 или по п. 12; и
инкубирование клеток.
14. Способ по п. 13, в котором инкубация клеток включает в себя пассирование клеток, причем выход продукции AAV в пассаже 7 является по меньшей мере в 2 раза большим по сравнению с контрольной линией клеток насекомых, содержащей бакуловирусный вектор, содержащий кассету экспрессии AAV Сар и кассету экспрессии AAV Rep, но не содержащий hr бакуловируса.
15. Способ по п. 13, в котором в пассаже 7 титр бакуловируса, содержащего кассету экспрессии AAV Сар, составляет более 21,5% от общего титра бакуловируса.
16. Способ выращивания AAV in vitro, включающий:
обеспечение культуры клеток насекомых;
инфицирование или трансфекцию клеток насекомых бакуловирусным вектором по любому из пп. 1-10; и
инкубирование клеток.
17. Способ по п. 16, в котором в Р7 выход AAV из клеток насекомых по меньшей мере на 50% превышает выход AAV в Р7 из клеток насекомых, содержащих бакуловирусный вектор без hr.
18. Способ по п. 16, в котором в Р7 выход AAV из клеток, содержащих hr бакуловируса, по меньшей мере на 20% превышает выход AAV из клеток, содержащих бакуловирусный вектор без hr.
19. Способ по п. 16, в котором бакуловирусный вектор не содержит Rep-связывающий элемент (RBE).
20. Способ продуцирования AAV in vitro, включающий выращивание культуры клеток насекомых, содержащей вектор по любому из пп. 1-10, и вектор, содержащий трансген, фланкированный AAV ITR.
21. Бакуловирусный вектор без Rep-связывающего элемента (RBE) для продуцирования AAV в клетках насекомых in vitro, содержащий:
кассету экспрессии AAV Сар;
кассету экспрессии AAV Rep; и
гомологичную область (hr) бакуловируса, расположенную примерно на расстоянии до около 4 тыс. п.о. от стартового кодона кассеты экспрессии AAV.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
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US201662325817P | 2016-04-21 | 2016-04-21 | |
US62/325,817 | 2016-04-21 | ||
PCT/US2017/028660 WO2017184879A1 (en) | 2016-04-21 | 2017-04-20 | Aav production in insect cells, methods and compositions therefor |
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RU2018128968A true RU2018128968A (ru) | 2020-05-21 |
RU2018128968A3 RU2018128968A3 (ru) | 2020-07-21 |
RU2739596C2 RU2739596C2 (ru) | 2020-12-28 |
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US (1) | US11414676B2 (ru) |
EP (1) | EP3445381A4 (ru) |
JP (2) | JP2019514352A (ru) |
KR (2) | KR20180134846A (ru) |
CN (1) | CN108699567B (ru) |
AU (1) | AU2017254652B2 (ru) |
CA (1) | CA3012699C (ru) |
IL (1) | IL261665B2 (ru) |
RU (1) | RU2739596C2 (ru) |
WO (1) | WO2017184879A1 (ru) |
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WO2016094783A1 (en) | 2014-12-12 | 2016-06-16 | Voyager Therapeutics, Inc. | Compositions and methods for the production of scaav |
EP3355938B1 (en) | 2015-10-01 | 2024-02-07 | Goleini Inc. | Targeted expression of chloride channels and methods of use thereof |
CA3024448A1 (en) | 2016-05-18 | 2017-11-23 | Voyager Therapeutics, Inc. | Modulatory polynucleotides |
JOP20190269A1 (ar) | 2017-06-15 | 2019-11-20 | Voyager Therapeutics Inc | بولي نوكليوتيدات aadc لعلاج مرض باركنسون |
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IL300407A (en) | 2020-08-10 | 2023-04-01 | Prevail Therapeutics Inc | Gene therapies for lysosomal disorders |
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CN1570121A (zh) * | 2003-07-25 | 2005-01-26 | 北京三诺佳邑生物技术有限责任公司 | 高效生产重组腺病毒伴随病毒的病毒载体、细胞系、方法及其组成的生产系统 |
CN101405033A (zh) * | 2006-01-20 | 2009-04-08 | 北卡罗来纳大学教堂山分校 | 增强感染性细小病毒载体在昆虫细胞中的生产 |
CA2655957C (en) | 2006-06-21 | 2016-05-03 | Amsterdam Molecular Therapeutics (Amt) B.V. | Vectors with modified initiation codon for the translation of aav-rep78 useful for production of aav in insect cells |
ES2385679T3 (es) * | 2006-08-24 | 2012-07-30 | Virovek, Inc. | Expresión de células de insecto de genes con marcos de lectura abiertos superpuestos, métodos y composiciones de éstos |
WO2009014445A2 (en) | 2007-07-26 | 2009-01-29 | Amsterdam Molecular Therapeutics B.V. | Baculoviral vectors comprising repeated coding sequences with differential codon biases |
ES2929031T3 (es) * | 2008-02-19 | 2022-11-24 | Uniqure Ip Bv | Optimización de la expresión de las proteínas rep y cap parvovirales en células de insecto |
US8679837B2 (en) * | 2009-04-02 | 2014-03-25 | University Of Florida Research Foundation, Inc. | Inducible system for highly efficient production of recombinant Adeno-associated virus (rAAV) vectors |
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2017
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- 2017-04-20 JP JP2018549544A patent/JP2019514352A/ja active Pending
- 2017-04-20 CN CN201780010098.0A patent/CN108699567B/zh active Active
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2018
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RU2739596C2 (ru) | 2020-12-28 |
IL261665B1 (en) | 2023-07-01 |
CN108699567A (zh) | 2018-10-23 |
AU2017254652B2 (en) | 2022-12-01 |
AU2017254652A1 (en) | 2018-08-09 |
WO2017184879A1 (en) | 2017-10-26 |
CA3012699A1 (en) | 2017-10-26 |
JP2024016212A (ja) | 2024-02-06 |
CN108699567B (zh) | 2024-02-13 |
EP3445381A1 (en) | 2019-02-27 |
KR20180134846A (ko) | 2018-12-19 |
KR20230079511A (ko) | 2023-06-07 |
EP3445381A4 (en) | 2019-10-02 |
IL261665B2 (en) | 2023-11-01 |
IL261665A (en) | 2018-10-31 |
RU2018128968A3 (ru) | 2020-07-21 |
US20180371495A1 (en) | 2018-12-27 |
CA3012699C (en) | 2023-11-21 |
US11414676B2 (en) | 2022-08-16 |
JP2019514352A (ja) | 2019-06-06 |
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