RU2560437C2 - Способ получения полимеров из белков шелка пауков - Google Patents
Способ получения полимеров из белков шелка пауков Download PDFInfo
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- RU2560437C2 RU2560437C2 RU2011145429/10A RU2011145429A RU2560437C2 RU 2560437 C2 RU2560437 C2 RU 2560437C2 RU 2011145429/10 A RU2011145429/10 A RU 2011145429/10A RU 2011145429 A RU2011145429 A RU 2011145429A RU 2560437 C2 RU2560437 C2 RU 2560437C2
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- Prior art keywords
- amino acid
- spider silk
- fragment
- acid residues
- silk protein
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43513—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae
- C07K14/43518—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from arachnidae from spiders
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Applications Claiming Priority (7)
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Families Citing this family (30)
| Publication number | Priority date | Publication date | Assignee | Title |
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| EP2547771B8 (en) * | 2010-03-18 | 2017-07-19 | Spiber Technologies AB | Production of proteins and polypeptides |
| DK3178843T3 (da) | 2010-10-27 | 2019-08-19 | Spiber Tech Ab | Edderkoppesilke-fusionsproteinstrukturer til at binde til et organisk mål |
| EP2716798B1 (en) * | 2011-06-01 | 2019-12-18 | Spiber Inc. | Artificial polypeptide fiber and method for producing same |
| JP6412866B2 (ja) * | 2012-05-02 | 2018-10-24 | スパイバー テクノロジーズ アーベーＳｐｉｂｅｒ Ｔｅｃｈｎｏｌｏｇｉｅｓ Ａｂ | 有機ターゲットに結合するための反復断片を持たないクモ糸融合タンパク質の構造 |
| ES2887366T3 (es) | 2013-09-17 | 2021-12-22 | Bolt Threads Inc | Métodos y composiciones para sintetizar fibras de seda mejoradas |
| PE20171791A1 (es) | 2014-12-02 | 2017-12-28 | Silk Therapeutics Inc | Prendas de vestir de seda y productos de alto rendimiento y metodos para elaborarla |
| CN114000218A (zh) * | 2015-03-16 | 2022-02-01 | 保尔特纺织品公司 | 改善的丝纤维 |
| EP3281948B1 (en) | 2015-04-09 | 2020-06-10 | Spiber Inc. | Polar solvent solution and production method thereof |
| JP6856828B2 (ja) | 2015-04-09 | 2021-04-14 | Ｓｐｉｂｅｒ株式会社 | 極性溶媒溶液及びその製造方法 |
| JP2018521239A (ja) * | 2015-06-11 | 2018-08-02 | ボルト スレッズ インコーポレイテッド | 改善された特性を有する組換えタンパク質繊維糸 |
| CN105031723B (zh) * | 2015-06-23 | 2017-05-24 | 上海交通大学 | 基于蜘蛛丝蛋白的温敏性水凝胶 |
| JP6956066B2 (ja) | 2015-07-14 | 2021-10-27 | エボルブド バイ ネイチャー， インコーポレイテッド | シルク性能衣服及び製品、並びにこれらを製造する方法 |
| JP2018159137A (ja) | 2015-08-20 | 2018-10-11 | 国立研究開発法人理化学研究所 | クモ糸様構造を有するポリペプチド繊維の製造方法 |
| EP3374378B1 (en) | 2015-11-13 | 2021-02-17 | Spiber Technologies AB | Charge-reversed n-terminal spider silk protein domain and uses thereof |
| GB201522610D0 (en) * | 2015-12-22 | 2016-02-03 | Univ Southampton | Protein |
| JP7168454B2 (ja) * | 2016-05-16 | 2022-11-09 | スピベル テクノロジーズ アクティエボラーグ | 固体表面のスパイダーシルクコーティング |
| JP2019518882A (ja) * | 2016-06-10 | 2019-07-04 | ボルト スレッズ インコーポレイテッド | 改善された特性を有する組換えタンパク質繊維糸 |
| EP3263593A1 (en) | 2016-07-01 | 2018-01-03 | Anna Rising | Engineered spider silk proteins and uses thereof |
| CA3035839A1 (en) | 2016-09-14 | 2018-03-22 | Bolt Threads, Inc. | Long uniform recombinant protein fibers |
| US20190275193A1 (en) * | 2016-11-11 | 2019-09-12 | Amsilk Gmbh | Use of a shrinkable biopolymer fiber as a sensor |
| CN107475807A (zh) * | 2017-08-30 | 2017-12-15 | 常州豫春化工有限公司 | 一种改性尼龙6纤维的制备方法 |
| EP3688017A1 (en) * | 2017-09-26 | 2020-08-05 | Spiber Technologies AB | Structuring of surface-active macromolecules |
| WO2019067745A1 (en) | 2017-09-27 | 2019-04-04 | Silk, Inc. | SILK-COATED FABRICS, PRODUCTS AND PREPARATION METHODS THEREFOR |
| JP2021035330A (ja) * | 2017-09-29 | 2021-03-04 | Ｓｐｉｂｅｒ株式会社 | 発現カセット |
| KR20220029660A (ko) * | 2019-07-04 | 2022-03-08 | 시비스 머테리얼 사이언스 리미티드 | 원핵생물 발현 시스템 및 이의 사용 방법 |
| CN110551194B (zh) * | 2019-09-19 | 2023-03-21 | 天津大学 | 重组蜘蛛卵鞘丝蛋白复合物及其生成的人造卵鞘丝 |
| EP4139337A4 (en) | 2020-04-23 | 2025-05-07 | Seevix Material Sciences Ltd. | COSMETIC COMPOSITIONS CONTAINING SPIDER SILK FROM DRAGLINE |
| WO2024091831A2 (en) * | 2022-10-28 | 2024-05-02 | National Taiwan University | A recombinant spider silk protein and method for tag-free and time-saving purification thereof |
| CN218413490U (zh) | 2022-10-31 | 2023-01-31 | 徐州龙安电子科技有限公司 | 一种新型蜘蛛电子标签 |
| DE102023105437B3 (de) | 2023-03-06 | 2024-06-13 | Dr. Ing. H.C. F. Porsche Aktiengesellschaft | Fahrzeugkomponente mit wenigstens einer Fahrzeuginnenraumkomponente |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007078239A2 (en) * | 2005-12-30 | 2007-07-12 | Spiber Technologies Ab | Spider silk proteins and methods for producing spider silk proteins |
| RU2007102846A (ru) * | 2004-07-22 | 2008-08-27 | Текнише Универзитет Мюнхен (DE) | Рекомбинантные белки шелка пауков |
| WO2008154547A2 (en) * | 2007-06-11 | 2008-12-18 | The Regents Of The University Of California | Spider silk dragline polynucleotides, polypeptides and methods of use thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2003201513A1 (en) * | 2002-01-11 | 2003-07-24 | Nexia Biotechnologies, Inc. | Methods of producing silk polypeptides and products thereof |
| BRPI0701826B1 (pt) * | 2007-03-16 | 2021-02-17 | Embrapa - Empresa Brasileira De Pesquisa Agropecuária | proteínas da teia de aranha nephilengys cruentata, avicularia juruensis e parawixia bistriata isoladas da biodiversidade brasileira |
-
2010
- 2010-04-21 US US13/264,938 patent/US8642734B2/en active Active
- 2010-04-21 EP EP10767397.2A patent/EP2421890B1/en active Active
- 2010-04-21 WO PCT/SE2010/050439 patent/WO2010123450A1/en not_active Ceased
- 2010-04-21 CA CA2759465A patent/CA2759465C/en not_active Expired - Fee Related
- 2010-04-21 AU AU2010239773A patent/AU2010239773B2/en not_active Ceased
- 2010-04-21 RU RU2011145429/10A patent/RU2560437C2/ru active
- 2010-04-21 CN CN2010800169433A patent/CN102395601A/zh active Pending
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- 2010-04-21 JP JP2012507182A patent/JP5912077B2/ja active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2007102846A (ru) * | 2004-07-22 | 2008-08-27 | Текнише Универзитет Мюнхен (DE) | Рекомбинантные белки шелка пауков |
| WO2007078239A2 (en) * | 2005-12-30 | 2007-07-12 | Spiber Technologies Ab | Spider silk proteins and methods for producing spider silk proteins |
| WO2008154547A2 (en) * | 2007-06-11 | 2008-12-18 | The Regents Of The University Of California | Spider silk dragline polynucleotides, polypeptides and methods of use thereof |
Non-Patent Citations (2)
| Title |
|---|
| DB "UniprotKB", N Q05H60_9ARAC, 14.11.2006. DB "EMBL-EBI", N * |
| HEDHAMMAR M. et al, Structural properties of recombinant nonrepetitive and repetitive parts of major ampullate spidroin 1 from Euprosthenops australis: implications for fiber formation, Biochemistry, 2008, v. 47, p. 3407-3417. * |
Also Published As
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| WO2010123450A1 (en) | 2010-10-28 |
| DK2421890T3 (en) | 2015-11-23 |
| US20120041177A1 (en) | 2012-02-16 |
| RU2011145429A (ru) | 2013-05-27 |
| CA2759465A1 (en) | 2010-10-28 |
| US8642734B2 (en) | 2014-02-04 |
| EP2421890A1 (en) | 2012-02-29 |
| EP2421890A4 (en) | 2012-10-24 |
| AU2010239773A1 (en) | 2011-11-03 |
| CA2759465C (en) | 2017-12-12 |
| JP5912077B2 (ja) | 2016-04-27 |
| AU2010239773B2 (en) | 2014-11-06 |
| JP2012531889A (ja) | 2012-12-13 |
| EP2421890B1 (en) | 2015-08-26 |
| CN102395601A (zh) | 2012-03-28 |
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