AR124480A1 - Métodos de purificación de proteínas de fusión con protección de carga - Google Patents
Métodos de purificación de proteínas de fusión con protección de cargaInfo
- Publication number
- AR124480A1 AR124480A1 ARP210103633A ARP210103633A AR124480A1 AR 124480 A1 AR124480 A1 AR 124480A1 AR P210103633 A ARP210103633 A AR P210103633A AR P210103633 A ARP210103633 A AR P210103633A AR 124480 A1 AR124480 A1 AR 124480A1
- Authority
- AR
- Argentina
- Prior art keywords
- protected
- fusion protein
- cargo
- biologically active
- cell lysate
- Prior art date
Links
- 108020001507 fusion proteins Proteins 0.000 title abstract 15
- 238000000034 method Methods 0.000 title abstract 10
- 238000001742 protein purification Methods 0.000 title 1
- 102000037865 fusion proteins Human genes 0.000 abstract 14
- 239000013592 cell lysate Substances 0.000 abstract 7
- 102000004169 proteins and genes Human genes 0.000 abstract 7
- 108090000623 proteins and genes Proteins 0.000 abstract 7
- 238000004191 hydrophobic interaction chromatography Methods 0.000 abstract 6
- 238000011210 chromatographic step Methods 0.000 abstract 4
- 238000005571 anion exchange chromatography Methods 0.000 abstract 2
- 238000005277 cation exchange chromatography Methods 0.000 abstract 2
- 238000012258 culturing Methods 0.000 abstract 2
- 238000010828 elution Methods 0.000 abstract 2
- 102000039446 nucleic acids Human genes 0.000 abstract 2
- 108020004707 nucleic acids Proteins 0.000 abstract 2
- 150000007523 nucleic acids Chemical class 0.000 abstract 2
- 238000004587 chromatography analysis Methods 0.000 abstract 1
- 230000002209 hydrophobic effect Effects 0.000 abstract 1
- 230000003993 interaction Effects 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/20—Partition-, reverse-phase or hydrophobic interaction chromatography
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
- A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/32—Bonded phase chromatography
- B01D15/325—Reversed phase
- B01D15/327—Reversed phase with hydrophobic interaction
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/36—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction
- B01D15/361—Ion-exchange
- B01D15/362—Cation-exchange
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/36—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction
- B01D15/361—Ion-exchange
- B01D15/363—Anion-exchange
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/42—Selective adsorption, e.g. chromatography characterised by the development mode, e.g. by displacement or by elution
- B01D15/424—Elution mode
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/18—Ion-exchange chromatography
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
-
- 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
- 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
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/62—DNA sequences coding for fusion proteins
-
- 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
- C12N9/14—Hydrolases (3)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
- C12N9/80—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)
- C12N9/82—Asparaginase (3.5.1.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y305/00—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
- C12Y305/01—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in linear amides (3.5.1)
- C12Y305/01001—Asparaginase (3.5.1.1)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/31—Fusion polypeptide fusions, other than Fc, for prolonged plasma life, e.g. albumin
-
- 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
- C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
Abstract
La presente invención se refiere a un método para purificar las proteínas con protección de carga a partir del lisado celular o liberado periplásmico usando cromatografía de interacción hidrofóbica como una primera etapa de cromatografía. Asimismo, en la presente, también se proveen las composiciones que comprenden proteínas con protección de carga y los métodos de tratamiento utilizando proteínas purificadas con protección de carga. Reivindicación 1: En algunas formas de realización, en la presente se proporciona un método para purificar una proteína de fusión con protección de carga a partir de un lisado celular o un liberado periplásmico, en donde la proteína de fusión con protección de carga comprende un dominio biológicamente activo y un dominio con protección de carga, y en donde el método comprende una cromatografía de interacción hidrofóbica como una primera etapa de la cromatografía. Reivindicación 2: En algunas formas de realización, en la presente se proporciona un método para producir una proteína de fusión con protección de carga a partir de un lisado celular o un liberado periplásmico, en donde la proteína de fusión con protección de carga comprende un dominio biológicamente activo y un dominio con protección de carga, en donde el método comprende i) cultivar células que comprenden un ácido nucleico que codifica la proteína de fusión con protección de carga; y ii) purificar la proteína de fusión con protección de carga, en donde la proteína con protección de carga se purifica a partir del lisado celular o liberado periplásmico usando cromatografía de interacción hidrofóbica como primera etapa de cromatografía. Reivindicación 17: Un método para producir una proteína de fusión PASilada biológicamente activa a partir de un lisado celular o un liberado periplásmico que comprende i) cultivar células que comprenden un ácido nucleico que codifica la proteína de fusión PASilada biológicamente activa; y ii) purificar la proteína de fusión PASilada biológicamente activa, y en donde la proteína PASilada biológicamente activa se purifica a partir del lisado celular o liberado periplásmico usando cromatografía de interacción hidrofóbica como primera etapa de cromatografía. Reivindicación 18: Un método para purificar una proteína de fusión con protección de carga que comprende un dominio biológicamente activo y un dominio de protección de carga de un lisado celular o liberación periplásmica, el método comprende las siguientes etapas en orden i) aplicar una solución de carga que comprende la proteína de fusión con protección de carga a una columna de cromatografía de interacción hidrofóbica; ii) aplicar una solución de lavado a la columna de cromatografía de interacción hidrofóbica; iii) aplicar una solución de elución a la columna de interacción hidrofóbica para eluir la proteína con protección de carga; vi) aplicar la proteína de fusión con protección de carga eluida en iii) como una solución de carga a una columna de cromatografía de intercambio aniónico; v) eluir la proteína de fusión con protección de carga de la columna de cromatografía de intercambio aniónico; vi) aplicar la proteína de fusión con protección de carga eluida en vi) como una solución de carga a una columna de cromatografía de intercambio catiónico; vii) aplicar una solución de lavado a la columna de cromatografía de intercambio viii) aplicar una solución de elución a la columna de cromatografía de intercambio de cationes para eluir la proteína con protección de carga.
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US202063130295P | 2020-12-23 | 2020-12-23 |
Publications (1)
Publication Number | Publication Date |
---|---|
AR124480A1 true AR124480A1 (es) | 2023-03-29 |
Family
ID=80118858
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
ARP210103633A AR124480A1 (es) | 2020-12-23 | 2021-12-22 | Métodos de purificación de proteínas de fusión con protección de carga |
Country Status (11)
Country | Link |
---|---|
US (1) | US20220227805A1 (es) |
EP (1) | EP4267592A1 (es) |
JP (1) | JP2024500974A (es) |
KR (1) | KR20230125179A (es) |
CN (1) | CN116916966A (es) |
AR (1) | AR124480A1 (es) |
AU (1) | AU2021410080A1 (es) |
CA (1) | CA3179177A1 (es) |
IL (1) | IL303948A (es) |
TW (1) | TW202241922A (es) |
WO (1) | WO2022140783A1 (es) |
Family Cites Families (19)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4551433A (en) | 1981-05-18 | 1985-11-05 | Genentech, Inc. | Microbial hybrid promoters |
US4755465A (en) | 1983-04-25 | 1988-07-05 | Genentech, Inc. | Secretion of correctly processed human growth hormone in E. coli and Pseudomonas |
US5281532A (en) | 1983-07-27 | 1994-01-25 | Mycogen Corporation | Pseudomas hosts transformed with bacillus endotoxin genes |
US4695455A (en) | 1985-01-22 | 1987-09-22 | Mycogen Corporation | Cellular encapsulation of pesticides produced by expression of heterologous genes |
US4695462A (en) | 1985-06-28 | 1987-09-22 | Mycogen Corporation | Cellular encapsulation of biological pesticides |
US5128130A (en) | 1988-01-22 | 1992-07-07 | Mycogen Corporation | Hybrid Bacillus thuringiensis gene, plasmid and transformed Pseudomonas fluorescens |
US5055294A (en) | 1988-03-03 | 1991-10-08 | Mycogen Corporation | Chimeric bacillus thuringiensis crystal protein gene comprising hd-73 and berliner 1715 toxin genes, transformed and expressed in pseudomonas fluorescens |
US5169760A (en) | 1989-07-27 | 1992-12-08 | Mycogen Corporation | Method, vectors, and host cells for the control of expression of heterologous genes from lac operated promoters |
US6406632B1 (en) | 1998-04-03 | 2002-06-18 | Symyx Technologies, Inc. | Rapid characterization of polymers |
US7294513B2 (en) | 2002-07-24 | 2007-11-13 | Wyatt Technology Corporation | Method and apparatus for characterizing solutions of small particles |
US9453251B2 (en) | 2002-10-08 | 2016-09-27 | Pfenex Inc. | Expression of mammalian proteins in Pseudomonas fluorescens |
AU2007275535B2 (en) | 2006-06-30 | 2012-08-16 | Servier IP UK Limited | Recombinant host for producing L-asparaginase II |
CA2663047A1 (en) | 2006-09-06 | 2008-03-13 | Phase Bioscience, Inc. | Therapeutic elastin-like polypeptide (elp) fusion proteins |
EP2054521A4 (en) * | 2006-10-03 | 2012-12-19 | Novo Nordisk As | METHODS OF PURIFYING CONJUGATES OF POLYPEPTIDES |
WO2011003633A1 (en) | 2009-07-06 | 2011-01-13 | Alize Pharma Ii | Pegylated l-asparaginase |
WO2013130684A1 (en) * | 2012-02-27 | 2013-09-06 | Amunix Operating Inc. | Xten-folate conjugate compositions and methods of making same |
EP4180525A1 (en) * | 2017-06-21 | 2023-05-17 | Jazz Pharmaceuticals Ireland Limited | Modified l-asparaginase |
EP3418383A1 (en) * | 2017-06-21 | 2018-12-26 | XL-protein GmbH | Modified l-asparaginase |
CN111278852A (zh) | 2017-10-27 | 2020-06-12 | 菲尼克斯公司 | 重组欧氏杆菌天冬酰胺酶的生产方法 |
-
2021
- 2021-12-22 WO PCT/US2021/073076 patent/WO2022140783A1/en active Application Filing
- 2021-12-22 TW TW110148097A patent/TW202241922A/zh unknown
- 2021-12-22 CA CA3179177A patent/CA3179177A1/en active Pending
- 2021-12-22 IL IL303948A patent/IL303948A/en unknown
- 2021-12-22 EP EP21851656.5A patent/EP4267592A1/en active Pending
- 2021-12-22 AR ARP210103633A patent/AR124480A1/es unknown
- 2021-12-22 KR KR1020237018060A patent/KR20230125179A/ko unknown
- 2021-12-22 AU AU2021410080A patent/AU2021410080A1/en active Pending
- 2021-12-22 CN CN202180079613.7A patent/CN116916966A/zh active Pending
- 2021-12-22 US US17/559,978 patent/US20220227805A1/en active Pending
- 2021-12-22 JP JP2023538915A patent/JP2024500974A/ja active Pending
Also Published As
Publication number | Publication date |
---|---|
IL303948A (en) | 2023-08-01 |
CA3179177A1 (en) | 2022-06-30 |
TW202241922A (zh) | 2022-11-01 |
US20220227805A1 (en) | 2022-07-21 |
EP4267592A1 (en) | 2023-11-01 |
JP2024500974A (ja) | 2024-01-10 |
AU2021410080A1 (en) | 2023-06-22 |
KR20230125179A (ko) | 2023-08-29 |
WO2022140783A1 (en) | 2022-06-30 |
CN116916966A (zh) | 2023-10-20 |
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