SE2030083A1 - Novel combination of tis sequence, signal peptide sequence and nucleic acid sequence encoding a recombinant protein - Google Patents
Novel combination of tis sequence, signal peptide sequence and nucleic acid sequence encoding a recombinant proteinInfo
- Publication number
- SE2030083A1 SE2030083A1 SE2030083A SE2030083A SE2030083A1 SE 2030083 A1 SE2030083 A1 SE 2030083A1 SE 2030083 A SE2030083 A SE 2030083A SE 2030083 A SE2030083 A SE 2030083A SE 2030083 A1 SE2030083 A1 SE 2030083A1
- Authority
- SE
- Sweden
- Prior art keywords
- seq
- nucleic acid
- acid sequence
- antibody
- dna construct
- Prior art date
Links
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Classifications
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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
- 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
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
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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
- 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
- C12N15/625—DNA sequences coding for fusion proteins containing a sequence coding for a signal sequence
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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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
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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
- 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
- C12N15/67—General methods for enhancing the expression
-
- 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
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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
- C12N2510/00—Genetically modified cells
- C12N2510/02—Cells for production
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2840/00—Vectors comprising a special translation-regulating system
- C12N2840/10—Vectors comprising a special translation-regulating system regulates levels of translation
- C12N2840/105—Vectors comprising a special translation-regulating system regulates levels of translation enhancing translation
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- Wood Science & Technology (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
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- General Health & Medical Sciences (AREA)
- Plant Pathology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Claims (28)
1. DNA-konstrukt för att uttrycka ett rekombinant protein hos mammaliaceller, varvidDNA-konstruktet innefattar:- en nukleinsyrasekvens enligt SEQ ID NO 1, varvid nukleinsyrasekvensenenligt SEQ ID NO 1 är en TIS-sekvens;- en nukleinsyrasekvens som kodar för en signalpeptid; och- en nukleinsyrasekvens som kodar för ett rekombinant protein, företrädesvis enmonoklonal antikropp, mer föredraget en IgG4 monoklonal antikropp;och varvid nukleinsyrasekvensen enligt SEQ ID NO 1 innefattar:- ATG-startkodonet i nukleinsyrasekvensen som kodar för förstaaminosyraresten hos signalpeptiden;- de två första nukleotidema nedströms ATG-startkodonet inukleinsyrasekvensen som kodar för den andra aminosyraresten hos signalpeptiden.
2. DNA-konstrukt enligt krav 1, varvid TIS-sekvensen transkriberas till ett RNA-motiv som fungerar som proteintranslationsinitieringsstället hos ett mRNA-transskript.
3. DNA-konstrukt enligt krav 1 eller 2, varvid TIS-sekvensen är en Kozak-sekvens somtranskriberas till ett RNA-motiv som fungerar som proteintranslationsinitieringsstället hos ett mRNA-transskript.
4. DNA-konstrukt enligt något av kraven 1-3, varvid nukleinsyrasekvensen som kodar för en signalpeptid innefattar en nukleinsyrasekvens enligt SEQ ID NO 2.
5. DNA-konstrukt enligt något av kraven 1-4, varvid nukleinsyrasekvensen enligt SEQID NO 1 innefattar:- ATG-startkodonet i SEQ ID NO 2; och- de två första nukleotidema nedströms ATG-startkodonet inukleinsyrasekvensen enligt SEQ ID NO 2.
6. DNA-konstrukt enligt något av kraven 1-5, varvid nukleinsyrasekvensen som kodarför signalpeptiden är operativt kopplad till nukleinsyrasekvensen som kodar för rekombinanta proteinet.
7. Expressionsvektor innefattande DNA-konstruktet enligt något av kraven 1-6.
8. Expressionskassett innefattande DNA-konstruktet enligt något av kraven 1-6.
9. Värdcell innefattande DNA-konstruktet enligt något av kraven 1-6, varvid nämnda värdcell företrädesvis är en eukaryot cell.
10. Värdcell enligt krav 9, varvid nämnda värdcell är en mammaliacell.
11. RNA uttryckt av DNA-konstruktet enligt något av kraven 1-6.
12. Metod för att uttrycka ett rekombinant protein, innefattande stegen att: - klona en eller flera öppna läsramar som kodar för ett rekombinant protein, elleren eller flera polypeptidkedj or därav, in i ett eller flera DNA-konstrukt enligtnågot av kraven 1-6; och - transfektera de resulterande nukleinsyrasekvensema in i en värdcell, varvid värdcellen företrädesvis är en eukaryot cell, mer föredraget en mammaliacell.
13. Metod enligt krav 12, vidare innefattande steget att integrera den transfekterade nukleinsyrasekvensen in i värdcellens genom.
Priority Applications (11)
| Application Number | Priority Date | Filing Date | Title |
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| SE2030083A SE544000C2 (en) | 2020-03-17 | 2020-03-17 | Novel combination of tis sequence, signal peptide sequence and nucleic acid sequence encoding a recombinant protein |
| EP21771233.0A EP4121543A4 (en) | 2020-03-17 | 2021-03-17 | NOVEL COMBINATION OF TIS SEQUENCE AND SIGNAL PEPTIDE SEQUENCE TO EXPRESS RECOMBINANT PROTEIN |
| KR1020227035849A KR102520468B1 (ko) | 2020-03-17 | 2021-03-17 | 재조합 단백질을 발현시키기 위한 tis 서열 및 신호 펩티드 서열의 신규한 조합 |
| PCT/SE2021/050233 WO2021188034A1 (en) | 2020-03-17 | 2021-03-17 | Novel combination of tis sequence and signal peptide sequence for expressing a recombinant protein |
| US17/911,772 US20230175008A1 (en) | 2020-03-17 | 2021-03-17 | Novel combination of tis sequence and signal peptide sequence for expressing a recombinant protein |
| CA3171532A CA3171532A1 (en) | 2020-03-17 | 2021-03-17 | Novel combination of tis sequence and signal peptide sequence for expressing a recombinant protein |
| AU2021239744A AU2021239744B2 (en) | 2020-03-17 | 2021-03-17 | Novel combination of TIS sequence and signal peptide sequence for expressing a recombinant protein |
| KR1020237011670A KR102661211B1 (ko) | 2020-03-17 | 2021-03-17 | 재조합 단백질을 발현시키기 위한 tis 서열 및 신호 펩티드 서열의 신규한 조합 |
| CN202180021454.5A CN115298313B (zh) | 2020-03-17 | 2021-03-17 | 用于表达重组蛋白的tis序列和信号肽序列的新组合 |
| JP2022554809A JP7441389B2 (ja) | 2020-03-17 | 2021-03-17 | 組換えタンパク質を発現させるためのｔｉｓ配列およびシグナルペプチド配列の新規の組み合わせ |
| AU2023201976A AU2023201976B2 (en) | 2020-03-17 | 2023-03-31 | Novel combination of TIS sequence and signal peptide sequence for expressing a recombinant protein |
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Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090239263A1 (en) * | 2005-10-04 | 2009-09-24 | Ucb Pharma S.A. | Expression Control Using Antibody Expression Optimisation Sequences |
| US20140120580A1 (en) * | 2001-08-27 | 2014-05-01 | Genentech, Inc. | System for antibody expression and assembly |
| WO2014093894A2 (en) * | 2012-12-13 | 2014-06-19 | The Trustees Of The University Of Pennsylvania | Dna antibody constructs and method of using same |
| WO2016081384A1 (en) * | 2014-11-17 | 2016-05-26 | Genentech, Inc. | Combination therapy comprising ox40 binding agonists and pd-1 axis binding antagonists |
| US20170349666A1 (en) * | 2014-11-20 | 2017-12-07 | Hoffmann-La Roche Inc. | Combination therapy of t cell activating bispecific antigen binding molecules and pd-1 axis binding antagonists |
| EP3323826A1 (en) * | 2016-11-21 | 2018-05-23 | Danmarks Tekniske Universitet | Native chinese hamster ovary cell secretion signal peptides for production of recombinant polypeptides |
| WO2018223182A1 (en) * | 2017-06-05 | 2018-12-13 | The Council Of The Queensland Institute Of Medical Research | A combination of, or a bispecific binding molecule to, an immune checkpoint molecule antagonist and a rank-l (nf- kb ligand) antagonist for cancer therapy or prophylaxis and uses therefor. |
| WO2019073954A1 (ja) * | 2017-10-10 | 2019-04-18 | 味の素株式会社 | タンパク質の製造法 |
| WO2019079496A2 (en) * | 2017-10-18 | 2019-04-25 | Regenxbio, Inc. | FULLY HUMAN ANTIBODY-BASED THERAPEUTIC AGENTS HAVING POST-TRANSLATION MODIFICATION |
-
2020
- 2020-03-17 SE SE2030083A patent/SE544000C2/en unknown
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140120580A1 (en) * | 2001-08-27 | 2014-05-01 | Genentech, Inc. | System for antibody expression and assembly |
| US20090239263A1 (en) * | 2005-10-04 | 2009-09-24 | Ucb Pharma S.A. | Expression Control Using Antibody Expression Optimisation Sequences |
| WO2014093894A2 (en) * | 2012-12-13 | 2014-06-19 | The Trustees Of The University Of Pennsylvania | Dna antibody constructs and method of using same |
| WO2016081384A1 (en) * | 2014-11-17 | 2016-05-26 | Genentech, Inc. | Combination therapy comprising ox40 binding agonists and pd-1 axis binding antagonists |
| US20170349666A1 (en) * | 2014-11-20 | 2017-12-07 | Hoffmann-La Roche Inc. | Combination therapy of t cell activating bispecific antigen binding molecules and pd-1 axis binding antagonists |
| EP3323826A1 (en) * | 2016-11-21 | 2018-05-23 | Danmarks Tekniske Universitet | Native chinese hamster ovary cell secretion signal peptides for production of recombinant polypeptides |
| WO2018223182A1 (en) * | 2017-06-05 | 2018-12-13 | The Council Of The Queensland Institute Of Medical Research | A combination of, or a bispecific binding molecule to, an immune checkpoint molecule antagonist and a rank-l (nf- kb ligand) antagonist for cancer therapy or prophylaxis and uses therefor. |
| WO2019073954A1 (ja) * | 2017-10-10 | 2019-04-18 | 味の素株式会社 | タンパク質の製造法 |
| WO2019079496A2 (en) * | 2017-10-18 | 2019-04-25 | Regenxbio, Inc. | FULLY HUMAN ANTIBODY-BASED THERAPEUTIC AGENTS HAVING POST-TRANSLATION MODIFICATION |
Non-Patent Citations (4)
| Title |
|---|
| Petersen S.D. et al., 'Modular 5 '-UTR hexamers for context-independent tuning of protein expression in eukaryotes', Nucleic Acids Research, 2018, Vol. 46, e127, pp. 1-11 * |
| Supplement to: Petersen S.D. et al., 'Modular 5 '-UTR hexamers for context-independent tuning of protein expression in eukaryotes', Nucleic Acids Research, 2018, Vol. 46, e127, pp. 1-11 * |
| You M et al., 'Efficient mAB production in CHO cells with optimized signal peptide, codon and UTR', Applied Microbiology and Biotechnology, 2018, Vol. 102, pp. 5953-5964 * |
| Zhou Y. et al., 'Enhancing full-length antibody production by signal peptide engineering', Microbial Cell Factories, 2016, Vol. 15, article 47, pp. 1-11 * |
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