HRP20201351T1 - Postupci proizvodnje dvolančanih proteina u bakteriji - Google Patents
Postupci proizvodnje dvolančanih proteina u bakteriji Download PDFInfo
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- HRP20201351T1 HRP20201351T1 HRP20201351TT HRP20201351T HRP20201351T1 HR P20201351 T1 HRP20201351 T1 HR P20201351T1 HR P20201351T T HRP20201351T T HR P20201351TT HR P20201351 T HRP20201351 T HR P20201351T HR P20201351 T1 HRP20201351 T1 HR P20201351T1
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- Prior art keywords
- tcr
- chain
- host cell
- immtac
- translation unit
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- 238000000034 method Methods 0.000 title claims 16
- 102000004169 proteins and genes Human genes 0.000 title claims 8
- 108090000623 proteins and genes Proteins 0.000 title claims 8
- 241000894006 Bacteria Species 0.000 title 1
- 108091008874 T cell receptors Proteins 0.000 claims 25
- 102000016266 T-Cell Antigen Receptors Human genes 0.000 claims 25
- 241000588724 Escherichia coli Species 0.000 claims 11
- 210000004027 cell Anatomy 0.000 claims 11
- 230000018109 developmental process Effects 0.000 claims 5
- 108091033319 polynucleotide Proteins 0.000 claims 4
- 102000040430 polynucleotide Human genes 0.000 claims 4
- 239000002157 polynucleotide Substances 0.000 claims 4
- 102000008394 Immunoglobulin Fragments Human genes 0.000 claims 3
- 108010021625 Immunoglobulin Fragments Proteins 0.000 claims 3
- 230000003698 anagen phase Effects 0.000 claims 3
- 239000001963 growth medium Substances 0.000 claims 3
- 238000013518 transcription Methods 0.000 claims 3
- 230000035897 transcription Effects 0.000 claims 3
- 108091026890 Coding region Proteins 0.000 claims 2
- 101710106383 Disulfide bond formation protein B Proteins 0.000 claims 2
- 102000009658 Peptidylprolyl Isomerase Human genes 0.000 claims 2
- 108010020062 Peptidylprolyl Isomerase Proteins 0.000 claims 2
- 101710116318 Probable disulfide formation protein Proteins 0.000 claims 2
- 210000001744 T-lymphocyte Anatomy 0.000 claims 2
- 239000000427 antigen Substances 0.000 claims 2
- 108091007433 antigens Proteins 0.000 claims 2
- 102000036639 antigens Human genes 0.000 claims 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims 2
- 230000001939 inductive effect Effects 0.000 claims 2
- 229910052760 oxygen Inorganic materials 0.000 claims 2
- 239000001301 oxygen Substances 0.000 claims 2
- 230000014621 translational initiation Effects 0.000 claims 2
- 206010028980 Neoplasm Diseases 0.000 claims 1
- 229910019142 PO4 Inorganic materials 0.000 claims 1
- 230000005867 T cell response Effects 0.000 claims 1
- 238000009825 accumulation Methods 0.000 claims 1
- 230000003115 biocidal effect Effects 0.000 claims 1
- 201000011510 cancer Diseases 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 238000012258 culturing Methods 0.000 claims 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 claims 1
- 239000003550 marker Substances 0.000 claims 1
- 230000035772 mutation Effects 0.000 claims 1
- 210000001322 periplasm Anatomy 0.000 claims 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims 1
- 239000010452 phosphate Substances 0.000 claims 1
- 108020003175 receptors Proteins 0.000 claims 1
- 101150065732 tir gene Proteins 0.000 claims 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- 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
- C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
- C07K14/70503—Immunoglobulin superfamily
- C07K14/7051—T-cell receptor (TcR)-CD3 complex
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
-
- 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
- C07K16/22—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against growth factors ; against growth regulators
-
- 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
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
- C07K16/244—Interleukins [IL]
-
- 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
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
- C07K16/244—Interleukins [IL]
- C07K16/247—IL-4
-
- 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
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/2809—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against the T-cell receptor (TcR)-CD3 complex
-
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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
-
- 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
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
- C07K2317/622—Single chain antibody (scFv)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Claims (14)
1. Postupak proizvodnje imunološki mobiliziranog monoklonskog receptora
T-stanice (ImmTAC) protiv raka, koji obuhvaća receptor T-stanice (TCR) – TCR alfa lanca i TCR beta lanca, u E.coli-stanici domaćina, naznačen time, da postupak obuhvaća sljedeće:
(a) kultiviranje E.coli-stanice domaćina, u svrhu ekspresije TCR alfa lanca i TCR beta lanca od ImmTAC, u mediju za uzgoj pod uvjetima koji obuhvaćaju sljedeće:
fazu rasta koja sadrži razvojnu temperaturu i razvojnu brzinu mućkanja, i
fazu proizvodnje koja sadrži produkcijsku temperaturu i produkcijsku brzinu mućkanja,
pri čemu se nakon ekspresije TCR alfa lanca i TCR beta lanca savijaju i sklapaju u svrhu tvorbe biološki aktivnog ImmTAC u E.coli-stanici domaćina;
gdje E.coli-stanica domaćina obuhvaća polinukleotid koji sadrži sljedeće:
(1) prvu translacijsku jedinicu koja kodira TCR alfa lanca od ImmTAC;
(2) drugu translacijsku jedinicu koja kodira TCR beta lanca od ImmTAC;
(3) treću translacijsku jedinicu koja kodira peptidil-prolil izomerazu, gdje peptidil-prolil izomeraza je FkpA protein; i
(4) četvrtu translacijsku jedinicu koja kodira protein disulfidne oksidoreduktaze, gdje protein disulfidne oksidoreduktaze je DsbC protein;
pri čemu za vrijeme faze rasta, razvojna temperatura leži u rasponu od 30°C do 34°C, dok za vrijeme faze proizvodnje, produkcijska temperatura leži u rasponu od 25°C do 29°C, gdje je za vrijeme faze rasta, razvojna brzina mućkanja dovoljna da se postigne maksimalna brzina nakupljanja kisika stanice domaćina, u rasponu od 3,5 do 4,5 mmol/L/min, te gdje je za vrijeme faze proizvodnje, produkcijska brzina mućkanja dovoljna da se postigne brzina unosa kisika stanice domaćina, u rasponu od 1,0 do 3,0 mmol/L/min; i
(b) dobivanje biološki aktivne ImmTAC iz E.coli-stanice domaćina.
2. Postupak prema patentnom zahtjevu 1, naznačen time, da polinukleotid dodatno obuhvaća tri kopije promotora, pri čemu je prva kopija u operativnoj kombinaciji s prvom translacijskom jedinicom, druga kopija je u operativnoj kombinaciji s drugom translacijskom jedinicom, i treća kopija je u operativnoj kombinaciji s trećom translacijskom jedinicom, u svrhu pokretanja transkripcije prvog lanca, drugog lanca i FkpA proteina, opcionalno gdje je promotor takav, da se može inducirati, opcionalno gdje promotor koji se može inducirati, jest promotor induciran kroz IPTG te kao takav pokreće transkripciju TCR alfa lanca, TCR beta lanca i FkpA proteina u odsutnosti IPTG-indukcije, i gdje je opcionalno promotor koji se može inducirati, Pho promotor koji pokreće transkripciju TCR alfa lanca, TCR beta lanca i FkpA proteina, kada se fosfat u mediju za uzgoj iscrpio.
3. Postupak prema patentnom zahtjevu 1 ili zahtjevu 2, naznačen time, da polinukleotid dodatno obuhvaća selektivan marker a medij za uzgoj obuhvaća selekcijski agens koji se sastoji od zasebnog antibiotika, kako bi se prouzročilo, da E.coli-stanica domaćina zadrži polinukleotid.
4. Postupak prema bilo kojem od patentnih zahtjeva 1 do 3, naznačen time, da prva translacijska jedinica sadrži prvu translacijsku inicijativnu regiju (TIR) u operativnoj kombinaciji s regijom kodiranja TCR alfa lanca, i druga translacijska jedinica sadrži drugu translacijsku inicijativnu regiju (TIR) u operativnoj kombinaciji s regijom kodiranja TCR beta lanca, pri čemu relativna snaga translacije prve i druge TIR, iznosi od oko 1,0 do oko 3,0.
5. Postupak prema bilo kojem od patentnih zahtjeva 1 do 4, naznačen time, da FkpA protein je E.coli FkpA.
6. Postupak prema patentnom zahtjevu 1, naznačen time, da DsbC protein je E.coli DsbC.
7. Postupak prema patentnom zahtjevu 1, naznačen time, da E.coli-stanica domaćina je soj s mutacijom degpS210A.
8. Postupak prema patentnom zahtjevu 7, naznačen time, da E.coli-stanica domaćina je soj s jednim genotipom od W3110 ΔfhuA ΔphoA ilvG2096 (Valr) Δprc spr43H1 ΔdegP ΔmanA lacIQ ΔompT ΔmenE degpS210.
9. Postupak prema bilo kojem od patentnih zahtjeva 1 do 8, naznačen time, da TCR alfa lanca obuhvaća jednu varijabilnu domenu TCR alfa lanca i jednu konstantnu domenu TCR alfa lanca, i time, da TCR beta lanca obuhvaća jednu varijabilnu domenu TCR beta lanca i jednu konstantnu domenu TCR beta lanca.
10. Postupak prema bilo kojem od patentnih zahtjeva 1 do 9, naznačen time, da su dva lanca od ImmTAC međusobno povezana pomoću najmanje jedne disulfidne veze.
11. Postupak prema bilo kojem od patentnih zahtjeva 1 do 10, naznačen time, da ImmTAC dodatno obuhvaća jedan fragment protutijela koji se veže na T-stanicu i aktivira odgovor T-stanice, i opcionalno pritom taj fragment protutijela sadrži fragment protutijela anti-CD3 jednostrukog lanca.
12. Postupak prema bilo kojem od patentnih zahtjeva 1 do 11, naznačen time, da ImmTAC sadrži jedan TCR modificiran inženjeringom kako bi posjedovao povećani afinitet za antigen, u usporedbi s TCR afinitetom na antigen od jednog TCR koji nije bio podvrgnut inženjeringu.
13. Postupak prema bilo kojem od patentnih zahtjeva 1 do 12, naznačen time, da se ImmTAC ponovno dobiva iz periplazme od E.coli-stanice domaćina.
14. Postupak prema bilo kojem od patentnih zahtjeva 1 do 13, naznačen time, da je razvojna brzina mućkanja veća od produkcijske brzine mućkanja za 10% do 40%.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201462075798P | 2014-11-05 | 2014-11-05 | |
| EP15797531.9A EP3215525B1 (en) | 2014-11-05 | 2015-11-05 | Methods of producing two chain proteins in bacteria |
| PCT/US2015/059342 WO2016073794A1 (en) | 2014-11-05 | 2015-11-05 | Methods of producing two chain proteins in bacteria |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| HRP20201351T1 true HRP20201351T1 (hr) | 2020-11-27 |
Family
ID=54602036
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| HRP20201351TT HRP20201351T1 (hr) | 2014-11-05 | 2020-08-27 | Postupci proizvodnje dvolančanih proteina u bakteriji |
Country Status (15)
| Country | Link |
|---|---|
| US (3) | US10066002B2 (hr) |
| EP (2) | EP3753948A1 (hr) |
| JP (2) | JP6770966B2 (hr) |
| KR (1) | KR102544705B1 (hr) |
| CN (2) | CN113699203A (hr) |
| AU (2) | AU2015342964B2 (hr) |
| BR (1) | BR112017009262A2 (hr) |
| CA (1) | CA2966573A1 (hr) |
| ES (1) | ES2819256T3 (hr) |
| HR (1) | HRP20201351T1 (hr) |
| IL (2) | IL252028B (hr) |
| MX (1) | MX2017005925A (hr) |
| RU (2) | RU2020141422A (hr) |
| SI (1) | SI3215525T1 (hr) |
| WO (1) | WO2016073794A1 (hr) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2966558C (en) * | 2014-11-05 | 2024-03-12 | Genentech, Inc. | Methods of producing two chain proteins in bacteria |
| RU2020141422A (ru) | 2014-11-05 | 2021-01-13 | Дженентек, Инк. | Способы получения двуцепочечных белков в бактериях |
| LT3430037T (lt) * | 2016-03-16 | 2022-12-12 | Immatics Biotechnologies Gmbh | Transfekuotos t ląstelės ir t ląstelių receptoriai, skirti naudoti vėžio gydymui taikant imunoterapiją |
| DE102016115246C5 (de) * | 2016-08-17 | 2018-12-20 | Immatics Biotechnologies Gmbh | Neue t-zellrezeptoren und deren verwendung in immuntherapie |
| ZA201900664B (en) | 2016-08-17 | 2021-09-29 | Paul Ehrlich Strasse 15 Tuebingen 72076 Germany | T cell receptors and immune therapy using the same |
| DE102017114737A1 (de) * | 2017-06-30 | 2019-01-03 | Immatics Biotechnologies Gmbh | Neue T-Zellrezeptoren und deren Verwendung in Immuntherapie |
| JP7278261B2 (ja) | 2017-08-23 | 2023-05-19 | ノバルティス アーゲー | マルチコピー遺伝子タンパク質発現系 |
| AU2019375413A1 (en) * | 2018-11-05 | 2021-05-27 | Genentech, Inc. | Methods of producing two chain proteins in prokaryotic host cells |
| CN109371049A (zh) * | 2018-11-14 | 2019-02-22 | 天津大学 | 分子伴侣在促进单克隆抗体的形成和/或提高单克隆抗体的表达量中的应用 |
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