RU2018128784A - Способы получения антител с заданным профилем гликозилирования - Google Patents
Способы получения антител с заданным профилем гликозилирования Download PDFInfo
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- RU2018128784A RU2018128784A RU2018128784A RU2018128784A RU2018128784A RU 2018128784 A RU2018128784 A RU 2018128784A RU 2018128784 A RU2018128784 A RU 2018128784A RU 2018128784 A RU2018128784 A RU 2018128784A RU 2018128784 A RU2018128784 A RU 2018128784A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
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- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0681—Cells of the genital tract; Non-germinal cells from gonads
- C12N5/0682—Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells
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- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/10—Transferases (2.)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
- C07K2317/41—Glycosylation, sialylation, or fucosylation
-
- 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
- C12N2510/00—Genetically modified cells
- C12N2510/02—Cells for production
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Engineering & Computer Science (AREA)
- Immunology (AREA)
- Biophysics (AREA)
- Reproductive Health (AREA)
- Cell Biology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Enzymes And Modification Thereof (AREA)
Claims (23)
1. Способ, включающий экспрессию антитела из мутантной по способу гликозилирования клеточной линии СНО, кодирующей указанное антитело, где указанная клеточная линия СНО подвергнута мутации так, что N-гликаны в антителах, продуцируемых клеткой, характеризуются концевым сахаром, который представляет собой N-ацетилглюкозамин.
2. Способ по п. 1, где мутация предусматривает по меньшей мере отрицательную регуляцию переносчика UDP-Gal в аппарат Гольджи.
3. Способ по п. 2, где мутация дополнительно предусматривает отрицательную регуляцию по меньшей мере переносчика СМР-сиаловой кислоты в аппарат Гольджи, отрицательную регуляцию или обеспечение неправильной локализации GlcNAc-TV, положительную регуляцию GlcNAc-TIII и их комбинации.
4. Способ по любому из пп. 1-3, где концевые остатки сахара в N-гликанах представляют собой N-ацетилглюкозамин.
5. Способ по любому из пп. 1-4, где клеточная линия представляет собой производственную клеточную линию.
6. Способ по любому из пп. 1-5, где клеточная линия выбрана из группы, включающей Lec3.2.8, Lec4.8, Lec4A.8, Lec8 и Lec10.8.
7. Способ по любому из пп. 1-4, который дополнительно включает стадию обеспечения реакции субстрата, представляющего собой N-ацетилглюкозамин, в N-гликане в антителе, экспрессированном из мутантной клеточной линии СНО, с реакционноспособным сахаром в присутствии катализатора-фермента с добавлением указанного остатка реакционноспособного сахара к гликану.
8. Способ по п. 7, где фермент представляет собой трансферазу.
9. Способ по п. 8, где трансфераза представляет собой фермент GalT, в том числе его мутантный вариант, где одна или две аминокислоты заменены, удалены или добавлены.
10. Способ по п. 9, где фермент GalT представляет собой мутантную форму, выбранную из группы, включающей Y289L, Y289N, Y289I и R228K.
11. Способ по п. 10, где фермент GALT характеризуется мутацией Y289L.
12. Способ по п. 7, где реакционноспособный сахар содержит химическую функциональную группу, выбранную из кетона, алкинила, азида.
13. Способ по п. 12, где реакционноспособный сахар представляет собой производное галактозамина.
14. Способ по п. 13, где реакционноспособный сахар содержит остаток GalAz или Keto-Gal.
15. Способ по любому из пп. 7-14, где реакционноспособный сахар представляет собой ферментный субстрат UDP-GalX в форме для переноса, где UDP представляет собой уридин-дифосфат, Gal представляет собой остаток галактозы, и X представляет собой функциональную группу в виде альдегида, алкинила или азида.
16. Способ по любому из пп. 7-15, который включает дополнительную стадию конъюгирования полезной нагрузки с альдегидной, алкинильной или азидной функциональными группами в остатке галактозы, добавленном к гликану путем переноса, опосредованного ферментом.
17. Способ по п. 16, где полезная нагрузка выбрана из выбрана из группы, включающей токсин, молекулу лекарственного средства (такого как цитотоксическое средство), полимер, антитело или его связывающий фрагмент.
18. Способ по п. 17, где молекула лекарственного средства выбрана из таковой, предусматривающей майтанзиноид, например N2'-деацетил-N2'-(3-меркапто-1-оксопропил)-майтанзин (DM1), N2'-деацетил-N2'-(4-меркапто-1-оксопентил)-майтанзин (DM3) и N2'-деацетил-N2'-(4-метил-4-меркапто-1-оксопентил)-майтанзин (DM4).
19. Способ по п. 17, где полезная нагрузка представляет собой токсин.
20. Способ по п. 17, где полимер представляет собой природный полимер, например крахмал или альбумин, или синтетический полимер, такой как PEG.
21. Способ по любому из пп. 16-20, где используемые химические реакции конъюгирования представляют собой реакции клик-химии.
22. Способ по п. 21, где реакции клик-химии представляют собой химические реакции без использования меди.
23. Молекула, полученная или получаемая в соответствии с любым из пп. 1-22.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201662287472P | 2016-01-27 | 2016-01-27 | |
| US62/287,472 | 2016-01-27 | ||
| PCT/US2017/015005 WO2017132298A1 (en) | 2016-01-27 | 2017-01-26 | Methods for preparing antibodies with a defined glycosylation pattern |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| RU2018128784A true RU2018128784A (ru) | 2020-02-27 |
| RU2018128784A3 RU2018128784A3 (ru) | 2020-04-28 |
Family
ID=59398609
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| RU2018128784A RU2018128784A (ru) | 2016-01-27 | 2017-01-26 | Способы получения антител с заданным профилем гликозилирования |
Country Status (9)
| Country | Link |
|---|---|
| US (1) | US11414477B2 (ru) |
| EP (1) | EP3407914A4 (ru) |
| JP (1) | JP6991978B2 (ru) |
| CN (1) | CN108495653A (ru) |
| AU (1) | AU2017212484C1 (ru) |
| BR (1) | BR112018015259A2 (ru) |
| CA (1) | CA3011734A1 (ru) |
| RU (1) | RU2018128784A (ru) |
| WO (1) | WO2017132298A1 (ru) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111902162A (zh) * | 2018-03-30 | 2020-11-06 | 三井化学株式会社 | 双亚氨基生物素化合物的药物递送用的用途 |
| GB202116442D0 (en) * | 2021-11-15 | 2021-12-29 | Actome Gmbh | High-efficacy labeling of antibodies with oligonucleotide labels |
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2017
- 2017-01-26 RU RU2018128784A patent/RU2018128784A/ru not_active Application Discontinuation
- 2017-01-26 CN CN201780008141.XA patent/CN108495653A/zh active Pending
- 2017-01-26 BR BR112018015259A patent/BR112018015259A2/pt not_active IP Right Cessation
- 2017-01-26 JP JP2018538706A patent/JP6991978B2/ja active Active
- 2017-01-26 CA CA3011734A patent/CA3011734A1/en not_active Abandoned
- 2017-01-26 AU AU2017212484A patent/AU2017212484C1/en not_active Ceased
- 2017-01-26 US US16/072,982 patent/US11414477B2/en active Active
- 2017-01-26 WO PCT/US2017/015005 patent/WO2017132298A1/en active Application Filing
- 2017-01-26 EP EP17744858.6A patent/EP3407914A4/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| JP6991978B2 (ja) | 2022-02-03 |
| EP3407914A1 (en) | 2018-12-05 |
| AU2017212484A1 (en) | 2018-08-09 |
| CN108495653A (zh) | 2018-09-04 |
| WO2017132298A1 (en) | 2017-08-03 |
| US20210214419A1 (en) | 2021-07-15 |
| EP3407914A4 (en) | 2019-08-07 |
| JP2019506161A (ja) | 2019-03-07 |
| AU2017212484B2 (en) | 2020-04-16 |
| AU2017212484C1 (en) | 2020-11-05 |
| WO2017132298A8 (en) | 2018-07-12 |
| CA3011734A1 (en) | 2017-08-03 |
| BR112018015259A2 (pt) | 2018-12-18 |
| RU2018128784A3 (ru) | 2020-04-28 |
| US11414477B2 (en) | 2022-08-16 |
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