RU2017122029A - Новые способы опосредованного ферментами конъюгирования полипептидов с использованием сортазы - Google Patents
Новые способы опосредованного ферментами конъюгирования полипептидов с использованием сортазы Download PDFInfo
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- RU2017122029A RU2017122029A RU2017122029A RU2017122029A RU2017122029A RU 2017122029 A RU2017122029 A RU 2017122029A RU 2017122029 A RU2017122029 A RU 2017122029A RU 2017122029 A RU2017122029 A RU 2017122029A RU 2017122029 A RU2017122029 A RU 2017122029A
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- Russia
- Prior art keywords
- seq
- amino acid
- polypeptide
- acid sequence
- terminus
- Prior art date
Links
- 230000001268 conjugating effect Effects 0.000 title 1
- 108090000765 processed proteins & peptides Proteins 0.000 claims 12
- 229920001184 polypeptide Polymers 0.000 claims 11
- 102000004196 processed proteins & peptides Human genes 0.000 claims 11
- 238000000034 method Methods 0.000 claims 6
- 125000003275 alpha amino acid group Chemical group 0.000 claims 5
- 125000000539 amino acid group Chemical group 0.000 claims 5
- 230000021615 conjugation Effects 0.000 claims 2
- 230000002255 enzymatic effect Effects 0.000 claims 2
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 claims 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 claims 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims 1
- 101900206500 Staphylococcus aureus Sortase A Proteins 0.000 claims 1
- 235000004279 alanine Nutrition 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 claims 1
- 235000018417 cysteine Nutrition 0.000 claims 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims 1
- 230000000694 effects Effects 0.000 claims 1
- 238000011534 incubation Methods 0.000 claims 1
- 239000011541 reaction mixture Substances 0.000 claims 1
Classifications
-
- 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
- C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
-
- 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)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
-
- 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)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6472—Cysteine endopeptidases (3.4.22)
-
- 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
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/22—Cysteine endopeptidases (3.4.22)
- C12Y304/2207—Sortase A (3.4.22.70)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Enzymes And Modification Thereof (AREA)
Claims (11)
1. Способ получения продукта ферментативного конъюгирования двух полипептидов, включающий следующие стадии:
инкубирование
i) первого полипептида, содержащего аминокислотную последовательность LPXTG (SEQ ID NO: 20, где Х может быть любым аминокислотным остатком),
ii) второго полипептида, который содержит i) аланинильное соединение на его N-конце или ii) олигоаланин Am (m равен 2 (SEQ ID NO: 26) или 3 (SEQ ID NO: 27), или 4 (SEQ ID NO: 28), или 5 (SEQ ID NO: 29)), или iii) аминокислотный остаток цистеина с последующими одним-тремя аминокислотными остатками аланина на его N-конце,
iii) третьего полипептида с сортазной активностью, который получен из сортазы А Staphylococcus aureus, и
выделение конъюгата из реакционной смеси и, посредством этого, получение продукта ферментативного конъюгирования двух полипептидов.
2. Способ по п. 1, в котором первый полипептид содержит на его С-конце аминокислотную последовательность LPXTG (SEQ ID NO: 20, где Х может быть любым аминокислотным остатком).
3. Способ по п. 2, в котором первый полипептид содержит на его С-конце аминокислотную последовательность LPETG (SEQ ID NO: 30).
4. Способ по любому из пп. 1-3, в котором второй полипептид имеет на его N-конце олигоаланин SEQ ID NO: 26 или SEQ ID NO: 27.
5. Способ по любому из пп. 1-4, в котором первый полипептид и второй полипептид независимо друг от друга выбраны из вариабельного домена антитела, фрагмента Fab тяжелой цепи антитела, области Fc антитела, метки, пептида, содержащего аминокислотную последовательность LPXTG (SEQ ID NO: 20, где Х может быть любым аминокислотным остатком), линкера и группировки несортазного мотива.
6. Способ по любому из пп. 1-5, в котором третий полипептид имеет аминокислотную последовательность SEQ ID NO: 21.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| EP14198532.5 | 2014-12-17 | ||
| EP14198532 | 2014-12-17 | ||
| PCT/EP2015/079617 WO2016096741A1 (en) | 2014-12-17 | 2015-12-14 | Novel methods for enzyme mediated polypeptide conjugation using sortase |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| RU2017122029A true RU2017122029A (ru) | 2019-01-17 |
| RU2017122029A3 RU2017122029A3 (ru) | 2019-07-17 |
| RU2714963C2 RU2714963C2 (ru) | 2020-02-21 |
Family
ID=52338848
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| RU2017122029A RU2714963C2 (ru) | 2014-12-17 | 2015-12-14 | Новые способы опосредованного ферментами конъюгирования полипептидов с использованием сортазы |
Country Status (11)
| Country | Link |
|---|---|
| US (1) | US10190182B2 (ru) |
| EP (1) | EP3233905B1 (ru) |
| JP (1) | JP6618539B2 (ru) |
| KR (1) | KR20170094216A (ru) |
| CN (1) | CN107001447B (ru) |
| BR (1) | BR112017010775A2 (ru) |
| CA (1) | CA2966287A1 (ru) |
| MX (1) | MX381723B (ru) |
| RU (1) | RU2714963C2 (ru) |
| SG (1) | SG11201704625UA (ru) |
| WO (1) | WO2016096741A1 (ru) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110295157A (zh) * | 2016-08-25 | 2019-10-01 | 北京大学 | 一种金黄色葡萄球菌分选酶a高效突变体 |
| WO2022166913A1 (en) * | 2021-02-04 | 2022-08-11 | Westlake Therapeutics (Hangzhou) Co. Limited | Modified red blood cells and uses thereof for treating hyperuricemia and gout |
| CN118786207A (zh) * | 2022-01-12 | 2024-10-15 | 西湖生物医药科技(上海)有限公司 | 经修饰的细胞及其用途 |
| CN114487409B (zh) * | 2022-04-14 | 2022-07-19 | 启德医药科技（苏州）有限公司 | 一种转肽酶活性的检测方法及检测试剂盒 |
| CN116949004B (zh) * | 2023-07-13 | 2024-04-09 | 苏州源启生物科技有限公司 | 一种转肽酶Sortase A及其制备方法和应用 |
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| US4150149A (en) | 1976-11-29 | 1979-04-17 | Professional Staff Association Of The Los Angeles County Harbor General Hospital | Method and means for the early detection and diagnosis of certain types of cancers |
| US4275149A (en) | 1978-11-24 | 1981-06-23 | Syva Company | Macromolecular environment control in specific receptor assays |
| US4318980A (en) | 1978-04-10 | 1982-03-09 | Miles Laboratories, Inc. | Heterogenous specific binding assay employing a cycling reactant as label |
| US4361544A (en) | 1980-03-03 | 1982-11-30 | Goldenberg Milton David | Tumor localization and therapy with labeled antibodies specific to intracellular tumor-associated markers |
| US4444744A (en) | 1980-03-03 | 1984-04-24 | Goldenberg Milton David | Tumor localization and therapy with labeled antibodies to cell surface antigens |
| US5316757A (en) | 1984-10-18 | 1994-05-31 | Board Of Regents, The University Of Texas System | Synthesis of polyazamacrocycles with more than one type of side-chain chelating groups |
| US5342606A (en) | 1984-10-18 | 1994-08-30 | Board Of Regents, The University Of Texas System | Polyazamacrocyclic compounds for complexation of metal ions |
| US4737456A (en) | 1985-05-09 | 1988-04-12 | Syntex (U.S.A.) Inc. | Reducing interference in ligand-receptor binding assays |
| US5959177A (en) | 1989-10-27 | 1999-09-28 | The Scripps Research Institute | Transgenic plants expressing assembled secretory antibodies |
| JP3951062B2 (ja) | 1991-09-19 | 2007-08-01 | ジェネンテック・インコーポレーテッド | 少なくとも遊離のチオールとして存在するシステインを有する抗体フラグメントの大腸菌での発現、２官能性Ｆ（ａｂ’）２抗体の産生のための使用 |
| US5739294A (en) | 1991-12-10 | 1998-04-14 | The Dow Chemical Company | Bicyclopol yazamacrocyclophosphonic acid complexes for use as contrast agents |
| US5480990A (en) | 1991-12-10 | 1996-01-02 | The Dow Chemical Company | Bicyclopolyazamacrocyclocarboxylic acid complexes for use as contrast agents |
| US5428139A (en) | 1991-12-10 | 1995-06-27 | The Dow Chemical Company | Bicyclopolyazamacrocyclophosphonic acid complexes for use as radiopharmaceuticals |
| ZA932522B (en) | 1992-04-10 | 1993-12-20 | Res Dev Foundation | Immunotoxins directed against c-erbB-2(HER/neu) related surface antigens |
| PT752248E (pt) | 1992-11-13 | 2001-01-31 | Idec Pharma Corp | Aplicacao terapeutica de anticorpos quimericos e marcados radioactivamente contra antigenios de diferenciacao restrita de linfocitos b humanos para o tratamento do linfoma de celulas b |
| US5385893A (en) | 1993-05-06 | 1995-01-31 | The Dow Chemical Company | Tricyclopolyazamacrocyclophosphonic acids, complexes and derivatives thereof, for use as contrast agents |
| US5462725A (en) | 1993-05-06 | 1995-10-31 | The Dow Chemical Company | 2-pyridylmethylenepolyazamacrocyclophosphonic acids, complexes and derivatives thereof, for use as contrast agents |
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| JP2014525904A (ja) | 2011-06-28 | 2014-10-02 | ホワイトヘッド・インスティテュート・フォー・バイオメディカル・リサーチ | タンパク質連結用クリックケミストリーハンドルを設置するためのソルターゼの使用 |
| KR20150023889A (ko) | 2012-06-27 | 2015-03-05 | 에프. 호프만-라 로슈 아게 | 2 개 이상의 상이한 결합 단위를 함유하는 맞춤-제작된 고도로 선별적이고 다중-특이적인 표적화 단위의 선별 및 제조 방법 및 이의 용도 |
-
2015
- 2015-12-14 RU RU2017122029A patent/RU2714963C2/ru active
- 2015-12-14 CN CN201580068354.2A patent/CN107001447B/zh active Active
- 2015-12-14 EP EP15817140.5A patent/EP3233905B1/en active Active
- 2015-12-14 JP JP2017531742A patent/JP6618539B2/ja active Active
- 2015-12-14 KR KR1020177016141A patent/KR20170094216A/ko not_active Ceased
- 2015-12-14 WO PCT/EP2015/079617 patent/WO2016096741A1/en not_active Ceased
- 2015-12-14 BR BR112017010775A patent/BR112017010775A2/pt active Search and Examination
- 2015-12-14 MX MX2017007503A patent/MX381723B/es unknown
- 2015-12-14 SG SG11201704625UA patent/SG11201704625UA/en unknown
- 2015-12-14 CA CA2966287A patent/CA2966287A1/en active Pending
-
2017
- 2017-06-16 US US15/625,975 patent/US10190182B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| RU2714963C2 (ru) | 2020-02-21 |
| KR20170094216A (ko) | 2017-08-17 |
| CA2966287A1 (en) | 2016-06-23 |
| EP3233905A1 (en) | 2017-10-25 |
| JP2018500901A (ja) | 2018-01-18 |
| EP3233905B1 (en) | 2020-01-29 |
| CN107001447B (zh) | 2021-07-23 |
| US10190182B2 (en) | 2019-01-29 |
| US20170292166A1 (en) | 2017-10-12 |
| BR112017010775A2 (pt) | 2018-02-27 |
| JP6618539B2 (ja) | 2019-12-11 |
| WO2016096741A1 (en) | 2016-06-23 |
| RU2017122029A3 (ru) | 2019-07-17 |
| MX381723B (es) | 2025-03-13 |
| MX2017007503A (es) | 2017-10-04 |
| SG11201704625UA (en) | 2017-07-28 |
| CN107001447A (zh) | 2017-08-01 |
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