RU2016128367A - Улучшенные способы получения рекомбинантного полипептида - Google Patents
Улучшенные способы получения рекомбинантного полипептида Download PDFInfo
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- RU2016128367A RU2016128367A RU2016128367A RU2016128367A RU2016128367A RU 2016128367 A RU2016128367 A RU 2016128367A RU 2016128367 A RU2016128367 A RU 2016128367A RU 2016128367 A RU2016128367 A RU 2016128367A RU 2016128367 A RU2016128367 A RU 2016128367A
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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 20
- 230000007017 scission Effects 0.000 claims 20
- 108091005804 Peptidases Proteins 0.000 claims 6
- 239000004365 Protease Substances 0.000 claims 6
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims 6
- 108010002231 IgA-specific serine endopeptidase Proteins 0.000 claims 2
- 230000008499 blood brain barrier function Effects 0.000 claims 2
- 210000004027 cell Anatomy 0.000 claims 2
- 238000012258 culturing Methods 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
- 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
- 108700011066 PreScission Protease Proteins 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
- 230000000593 degrading effect Effects 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
- 229960004072 thrombin Drugs 0.000 claims 1
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/005—Glycopeptides, glycoproteins
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
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- C07K—PEPTIDES
- 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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- C07K—PEPTIDES
- 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
- C07K14/48—Nerve growth factor [NGF]
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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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- C07K16/00—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
- C07K16/2881—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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- C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
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- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- 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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- 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
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- C07K2317/00—Immunoglobulins specific features
- 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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- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
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- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
- C07K2317/522—CH1 domain
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- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
- C07K2317/524—CH2 domain
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- C07K2317/526—CH3 domain
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- C07K2317/53—Hinge
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- C07K2317/55—Fab or Fab'
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- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
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- C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
- C07K2317/622—Single chain antibody (scFv)
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- C07K2319/00—Fusion polypeptide
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- C07K2319/50—Fusion polypeptide containing protease site
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- C07K2319/60—Fusion polypeptide containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]
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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 IgA-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. IgG-degrading enzyme of Streptococcus pyogenes - фермент Streptococcus pyogenes, расщепляющий IgG), 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 IgA-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 (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP13198738 | 2013-12-20 | ||
| EP13198738.0 | 2013-12-20 | ||
| PCT/EP2014/077196 WO2015091144A1 (en) | 2013-12-20 | 2014-12-10 | Improved recombinant polypeptide production methods |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| RU2019134445A Division RU2019134445A (ru) | 2013-12-20 | 2014-12-10 | Улучшенные способы получения рекомбинантного полипептида |
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| Publication Number | Publication Date |
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| RU2016128367A true RU2016128367A (ru) | 2018-01-25 |
| RU2711322C1 RU2711322C1 (ru) | 2020-01-16 |
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| RU2016128367A RU2711322C1 (ru) | 2013-12-20 | 2014-12-10 | Улучшенные способы получения рекомбинантного полипептида |
| RU2019134445A RU2019134445A (ru) | 2013-12-20 | 2014-12-10 | Улучшенные способы получения рекомбинантного полипептида |
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| Country | Link |
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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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Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| HUE057952T2 (hu) | 2015-06-24 | 2022-06-28 | Hoffmann La Roche | Anti-transzferrin receptor antitestek testreszabott affinitással |
| JP6657392B2 (ja) | 2015-10-02 | 2020-03-04 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | 二重特異性抗ヒトｃｄ２０／ヒトトランスフェリン受容体抗体及び使用方法 |
| 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 |
| KR102709884B1 (ko) * | 2017-11-30 | 2024-09-26 | 리제너론 파마슈티칼스 인코포레이티드 | 인간화된 trkb 유전자좌를 포함하는 비인간 동물 |
| CN112105634B (zh) * | 2018-09-21 | 2024-03-12 | 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 |
| AU2021226352A1 (en) * | 2020-02-25 | 2022-09-29 | Momenta Pharmaceuticals, Inc. | Enzymes for sialylation of glycans |
| 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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| CA2862820A1 (en) * | 2012-02-29 | 2013-09-06 | F. Hoffmann-La Roche Ag | On-column enzymatic cleavage |
| EP2668901A1 (en) | 2012-05-31 | 2013-12-04 | Roche Diagniostics GmbH | Sensor insertion assembly, sensor cartridge, and inserter |
| TR201909156T4 (tr) | 2012-08-29 | 2019-07-22 | Hoffmann La Roche | Kan beyin bariyeri mekiği. |
-
2014
- 2014-12-10 RU RU2016128367A patent/RU2711322C1/ru not_active IP Right Cessation
- 2014-12-10 WO PCT/EP2014/077196 patent/WO2015091144A1/en active Application Filing
- 2014-12-10 RU RU2019134445A patent/RU2019134445A/ru unknown
- 2014-12-10 US US15/106,205 patent/US10370692B2/en active Active
- 2014-12-10 CN CN201480069054.1A patent/CN105829542A/zh active Pending
- 2014-12-10 CA CA2929149A patent/CA2929149A1/en not_active Abandoned
- 2014-12-10 KR KR1020167016420A patent/KR20160098277A/ko not_active Application Discontinuation
- 2014-12-10 MX MX2016007207A patent/MX368656B/es active IP Right Grant
- 2014-12-10 JP JP2016541174A patent/JP6618912B2/ja not_active Expired - Fee Related
- 2014-12-10 BR BR112016012229A patent/BR112016012229A2/pt not_active Application Discontinuation
- 2014-12-10 EP EP14827183.6A patent/EP3083977B1/en active Active
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2016
- 2016-10-25 HK HK16112252.0A patent/HK1223980A1/zh unknown
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Also Published As
| Publication number | Publication date |
|---|---|
| EP3083977B1 (en) | 2018-02-28 |
| RU2711322C1 (ru) | 2020-01-16 |
| EP3083977A1 (en) | 2016-10-26 |
| JP2017500040A (ja) | 2017-01-05 |
| US11098338B2 (en) | 2021-08-24 |
| KR20160098277A (ko) | 2016-08-18 |
| US20160326562A1 (en) | 2016-11-10 |
| MX368656B (es) | 2019-10-10 |
| US20190271021A1 (en) | 2019-09-05 |
| CN105829542A (zh) | 2016-08-03 |
| WO2015091144A1 (en) | 2015-06-25 |
| JP6618912B2 (ja) | 2019-12-11 |
| MX2016007207A (es) | 2016-07-21 |
| RU2019134445A (ru) | 2019-12-20 |
| US10370692B2 (en) | 2019-08-06 |
| HK1223980A1 (zh) | 2017-08-11 |
| BR112016012229A2 (pt) | 2017-09-26 |
| CA2929149A1 (en) | 2015-06-25 |
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| MM4A | The patent is invalid due to non-payment of fees |
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