RU2019134445A - Улучшенные способы получения рекомбинантного полипептида - Google Patents
Улучшенные способы получения рекомбинантного полипептида Download PDFInfo
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims 20
- 229920001184 polypeptide Polymers 0.000 title claims 18
- 102000004196 processed proteins & peptides Human genes 0.000 title claims 18
- 238000000034 method Methods 0.000 title claims 9
- 238000003776 cleavage reaction Methods 0.000 claims 19
- 230000007017 scission Effects 0.000 claims 19
- 108091005804 Peptidases Proteins 0.000 claims 8
- 239000004365 Protease Substances 0.000 claims 8
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims 8
- 230000008499 blood brain barrier function Effects 0.000 claims 2
- 210000004027 cell Anatomy 0.000 claims 2
- 238000012258 culturing Methods 0.000 claims 2
- 239000001963 growth medium Substances 0.000 claims 2
- 210000004962 mammalian cell Anatomy 0.000 claims 2
- 102000039446 nucleic acids Human genes 0.000 claims 2
- 108020004707 nucleic acids Proteins 0.000 claims 2
- 150000007523 nucleic acids Chemical class 0.000 claims 2
- 102000005600 Cathepsins Human genes 0.000 claims 1
- 108010084457 Cathepsins Proteins 0.000 claims 1
- 108010013369 Enteropeptidase Proteins 0.000 claims 1
- 102100029727 Enteropeptidase Human genes 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 claims 1
- 108090001126 Furin Proteins 0.000 claims 1
- 102000004961 Furin Human genes 0.000 claims 1
- 102000001398 Granzyme Human genes 0.000 claims 1
- 108060005986 Granzyme Proteins 0.000 claims 1
- 108010002231 IgA-specific serine endopeptidase Proteins 0.000 claims 1
- 102000005741 Metalloproteases Human genes 0.000 claims 1
- 108010006035 Metalloproteases Proteins 0.000 claims 1
- 108010025020 Nerve Growth Factor Proteins 0.000 claims 1
- 102000007072 Nerve Growth Factors Human genes 0.000 claims 1
- 241000193996 Streptococcus pyogenes Species 0.000 claims 1
- 108090000787 Subtilisin Proteins 0.000 claims 1
- 108010076818 TEV protease Proteins 0.000 claims 1
- 108090000190 Thrombin Proteins 0.000 claims 1
- 241000723792 Tobacco etch virus Species 0.000 claims 1
- 241000723873 Tobacco mosaic virus Species 0.000 claims 1
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 125000000539 amino acid group Chemical group 0.000 claims 1
- 210000001218 blood-brain barrier Anatomy 0.000 claims 1
- 230000002255 enzymatic effect Effects 0.000 claims 1
- 229940088598 enzyme Drugs 0.000 claims 1
- 239000003900 neurotrophic factor Substances 0.000 claims 1
- 229940012957 plasmin Drugs 0.000 claims 1
- 238000000746 purification Methods 0.000 claims 1
- 239000002510 pyrogen Substances 0.000 claims 1
- 229960004072 thrombin Drugs 0.000 claims 1
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- C12P21/00—Preparation of peptides or proteins
- C12P21/005—Glycopeptides, glycoproteins
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
- C07K14/48—Nerve growth factor [NGF]
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- C—CHEMISTRY; METALLURGY
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- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
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- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2881—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against CD71
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- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/10—Tetrapeptides
- C07K5/1019—Tetrapeptides with the first amino acid being basic
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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
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- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
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- C07K2317/522—CH1 domain
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- C07K2317/524—CH2 domain
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Claims (12)
1. Способ получения рекомбинантного полипептида с использованием вариантного прополипептида, включающий следующие этапы:
культивирование клетки млекопитающего, содержащей нуклеиновую кислоту, кодирующую полипептид в виде прополипептида (гибридного полипептида из просегмента и полипептида), в котором эндогенный сайт ферментативного расщепления между просегментом и полипептидом заменяется экзогенным сайтом расщепления протеазой,
извлечение рекомбинантного вариантного полипептида или вариантного прополипептида из клетки или культивационной среды и, таким образом, получение рекомбинантного полипептида.
2. Способ по п. 1, где экзогенный сайт расщепления протеазой выбирают из группы, содержащей сайт расщепления плазмином, сайт расщепления фурином, сайт расщепления lgA-протеазой, сайт расщепления протеазой TEV (от англ. tobacco etch virus - вирус табачной мозаики), сайт расщепления гранзимом В, сайт расщепления тромбином, сайт расщепления фактором 10, сайт расщепления энтерокиназой, сайт расщепления субтилизином, сайт расщепления катепсином, сайт расщепления металлопротеиназой, сайт расщепления протеазой IDES (от англ. lgG-degrading enzyme of Streptococcus pyogenes - фермент Streptococcus pyogenes, расщепляющий lgG), сайт расщепления протеазой PreScission или их функциональные варианты.
3. Способ по п. 1, где расщепление прополипептида осуществляют во время очистки полипептида.
4. Способ по любому из пп. 1-3, где экзогенная протеаза представляет собой lgA-протеазу.
5. Способ получения гибридного полипептида, включающий следующие этапы:
культивирование клетки млекопитающего, содержащей нуклеиновую кислоту, кодирующую вариантный гибридный полипептид, где аминокислотная последовательность гибридного полипептида была модифицирована путем замены в гибридном прополипептиде эндогенного сайта расщепления протеазой между просегментом и гибридным полипептидом на экзогенный (относительно происхождения частей полипептида) или искусственный сайт расщепления протеазой,
извлечение гибридного полипептида или гибридного прополипептида из клетки или культивационной среды и, таким образом, получение (рекомбинантного) гибридного полипептида.
6. Способ по п. 1 или 5, характеризующийся тем, что гибридный полипептид содержит биологически активную группу, линкерный пептид и одновалентную связывающую группу, которая связывается с рецептором гематоэнцефалического барьера (ВВВ, от англ. blood-brain-barier).
7. Способ по п. 6, характеризующийся тем, что линкерный пептид содержит один или более чем один отрицательно заряженный аминокислотный остаток.
8. Способ по п. 1 или 5, характеризующийся тем, что полипептид представляет собой нейротрофический фактор.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP13198738 | 2013-12-20 | ||
| EP13198738.0 | 2013-12-20 |
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| RU2016128367A Division RU2711322C1 (ru) | 2013-12-20 | 2014-12-10 | Улучшенные способы получения рекомбинантного полипептида |
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| RU2019134445A RU2019134445A (ru) | 2013-12-20 | 2014-12-10 | Улучшенные способы получения рекомбинантного полипептида |
| RU2016128367A RU2711322C1 (ru) | 2013-12-20 | 2014-12-10 | Улучшенные способы получения рекомбинантного полипептида |
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| US (2) | US10370692B2 (ru) |
| EP (1) | EP3083977B1 (ru) |
| JP (1) | JP6618912B2 (ru) |
| KR (1) | KR20160098277A (ru) |
| CN (1) | CN105829542A (ru) |
| BR (1) | BR112016012229A2 (ru) |
| CA (1) | CA2929149A1 (ru) |
| HK (1) | HK1223980A1 (ru) |
| MX (1) | MX368656B (ru) |
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| LT3313879T (lt) | 2015-06-24 | 2022-03-25 | F. Hoffmann-La Roche Ag | Antikūnai prieš transferino receptorių su pritaikytu giminingumu |
| TWI819458B (zh) | 2015-10-02 | 2023-10-21 | 瑞士商赫孚孟拉羅股份公司 | 雙特異性抗‐人類cd20/人類轉鐵蛋白受體抗體及使用方法 |
| AR106189A1 (es) | 2015-10-02 | 2017-12-20 | Hoffmann La Roche | ANTICUERPOS BIESPECÍFICOS CONTRA EL A-b HUMANO Y EL RECEPTOR DE TRANSFERRINA HUMANO Y MÉTODOS DE USO |
| SG11202002456WA (en) * | 2017-11-30 | 2020-04-29 | Regeneron Pharma | Non-human animals comprising a humanized trkb locus |
| WO2020057645A1 (zh) * | 2018-09-21 | 2020-03-26 | 信达生物制药(苏州)有限公司 | 新型白介素2及其用途 |
| KR102027955B1 (ko) | 2019-01-24 | 2019-10-02 | 이현종 | 로스터 후드용 내부커버 및 이의 제조방법 |
| BR112022016948A2 (pt) * | 2020-02-25 | 2022-11-22 | Momenta Pharmaceuticals Inc | Enzimas para sialilação de glicanos |
| IT202000032423A1 (it) * | 2020-12-24 | 2022-06-24 | Univ Degli Studi Di Trieste | Metodo per la produzione di forme processate proteoliticamente di fattori trofici o fattori di crescita |
| US20230340058A1 (en) * | 2021-11-11 | 2023-10-26 | Tectonic Therapeutic, Inc. | Relaxin-2 fusion protein analogs and methods of using same |
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- 2014-12-10 EP EP14827183.6A patent/EP3083977B1/en active Active
- 2014-12-10 US US15/106,205 patent/US10370692B2/en active Active
- 2014-12-10 BR BR112016012229A patent/BR112016012229A2/pt not_active Application Discontinuation
- 2014-12-10 CA CA2929149A patent/CA2929149A1/en not_active Abandoned
- 2014-12-10 CN CN201480069054.1A patent/CN105829542A/zh active Pending
- 2014-12-10 RU RU2019134445A patent/RU2019134445A/ru unknown
- 2014-12-10 WO PCT/EP2014/077196 patent/WO2015091144A1/en active Application Filing
- 2014-12-10 JP JP2016541174A patent/JP6618912B2/ja active Active
- 2014-12-10 RU RU2016128367A patent/RU2711322C1/ru not_active IP Right Cessation
- 2014-12-10 KR KR1020167016420A patent/KR20160098277A/ko not_active Application Discontinuation
-
2016
- 2016-10-25 HK HK16112252.0A patent/HK1223980A1/zh unknown
-
2019
- 2019-05-15 US US16/413,063 patent/US11098338B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| US20190271021A1 (en) | 2019-09-05 |
| CN105829542A (zh) | 2016-08-03 |
| US10370692B2 (en) | 2019-08-06 |
| HK1223980A1 (zh) | 2017-08-11 |
| US11098338B2 (en) | 2021-08-24 |
| MX2016007207A (es) | 2016-07-21 |
| WO2015091144A1 (en) | 2015-06-25 |
| EP3083977A1 (en) | 2016-10-26 |
| EP3083977B1 (en) | 2018-02-28 |
| CA2929149A1 (en) | 2015-06-25 |
| RU2711322C1 (ru) | 2020-01-16 |
| JP6618912B2 (ja) | 2019-12-11 |
| US20160326562A1 (en) | 2016-11-10 |
| JP2017500040A (ja) | 2017-01-05 |
| BR112016012229A2 (pt) | 2017-09-26 |
| KR20160098277A (ko) | 2016-08-18 |
| MX368656B (es) | 2019-10-10 |
| RU2016128367A (ru) | 2018-01-25 |
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