JP6599980B2 - タンパク質抽出方法 - Google Patents
タンパク質抽出方法 Download PDFInfo
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- JP6599980B2 JP6599980B2 JP2017515662A JP2017515662A JP6599980B2 JP 6599980 B2 JP6599980 B2 JP 6599980B2 JP 2017515662 A JP2017515662 A JP 2017515662A JP 2017515662 A JP2017515662 A JP 2017515662A JP 6599980 B2 JP6599980 B2 JP 6599980B2
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- JP
- Japan
- Prior art keywords
- column
- volume
- sample
- extraction
- histone
- Prior art date
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Classifications
-
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4702—Regulators; Modulating activity
-
- 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
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/12—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the preparation of the feed
-
- 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
- B01D15/34—Size selective separation, e.g. size exclusion chromatography, gel filtration, permeation
-
- 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
-
- 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
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4702—Regulators; Modulating activity
- C07K14/4703—Inhibitors; Suppressors
Description
本明細書の実施形態をより容易に理解できるように用語を定義する。追加の定義は、詳細な説明全体を通して記載されている。
Claims (10)
- DNA凝縮タンパク質をDNA凝縮タンパク質を含む調製物から抽出する方法であって、前記調製物は細胞培養上清、体液、組織ホモジネート、及び分画プロセスからの一画分からなる群から選択され、
(i)前記調製物を有機カチオン、中性塩及び非イオン界面活性剤とpH0.1から1.0で接触させるステップ、
(ii)混合物を30分間から150分間インキュベートするステップ、
(iii)固体を除去して抽出物を得るステップ、
(iv)前記抽出物を(1)アニオン交換クロマトグラフィを空隙排除モードで実施するのに適切な正電荷粒子又は(2)排除限界が5,000ダルトンの実質的に無電荷のサイズ排除クロマトグラフィ粒子を含む粒子充填カラムにかけるステップであって、前記粒子充填カラムは粒子間体積を有し、
前記カラムにかけた抽出物の体積は前記粒子間体積以下である、ステップ、
(v)前記ステップ(iv)の後に、緩衝剤を前記カラムに通して、試料を前記カラムの下方に移し、前記カラムの排除体積に対応するピークにおいて、抽出されたDNA凝縮タンパク質を収集するステップ
を含み、前記有機カチオンが、0.01%から0.1%(重量/体積)の濃度のエタクリジン若しくはメチレンブルー、又は0.5Mから1.5Mの濃度のグアニジニウムである、方法。 - 前記有機カチオンが、エタクリジンである、請求項1に記載の方法。
- エタクリジン又はメチレンブルーの濃度が、0.02から0.05%、又は0.03から0.10%(重量/体積)の範囲である、請求項1又は2のいずれか一項に記載の方法。
- グアニジニウムの濃度が、0.8から1.2M、又は0.9から1.1Mの範囲である、請求項1に記載の方法。
- 前記中性塩が1Mから3Mの濃度範囲で存在する、請求項1に記載の方法。
- 前記中性塩が塩化ナトリウム又は塩化カリウムである、請求項1に記載の方法。
- 前記非イオン界面活性剤が、Tween(登録商標)20、Tween(登録商標)40、Tween(登録商標)60及びノニデット(Nonidet)(登録商標)NP40からなる群から選択される、請求項1に記載の方法。
- 前記非イオン界面活性剤が0.05%から0.25%w/vの濃度範囲で存在する、請求項1に記載の方法。
- 前記DNA凝縮タンパク質が1種以上のヒストンタンパク質を含む、請求項1に記載の方法。
- 前記調製物が、細胞培養上清である、請求項1に記載の方法。
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/SG2014/000237 WO2015183183A1 (en) | 2014-05-29 | 2014-05-29 | Protein Extraction Methods |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2017519044A JP2017519044A (ja) | 2017-07-13 |
JP6599980B2 true JP6599980B2 (ja) | 2019-10-30 |
Family
ID=54699375
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2017515662A Expired - Fee Related JP6599980B2 (ja) | 2014-05-29 | 2014-05-29 | タンパク質抽出方法 |
Country Status (7)
Country | Link |
---|---|
US (1) | US10259851B2 (ja) |
EP (1) | EP3149023A4 (ja) |
JP (1) | JP6599980B2 (ja) |
KR (1) | KR20170010750A (ja) |
CN (1) | CN106232614A (ja) |
SG (1) | SG11201609815QA (ja) |
WO (1) | WO2015183183A1 (ja) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US11385238B2 (en) * | 2017-06-30 | 2022-07-12 | Amgen Inc. | Methods of protein clips recovery |
CN114072006A (zh) * | 2019-07-02 | 2022-02-18 | 罗盖特公司 | 不含痕量的有机溶剂的低脂质含量燕麦蛋白质组合物 |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SU1319352A1 (ru) * | 1985-01-07 | 1995-09-10 | Ленинградский государственный университет им.А.А.Жданова | Способ очистки препарата гистона н4 из ткани тимуса |
CN1053195C (zh) * | 1997-08-04 | 2000-06-07 | 李京华 | 从鸡血中提取鸡核酸及核蛋白的方法 |
CN1233654C (zh) * | 2004-05-10 | 2005-12-28 | 四川大学 | 羟基磷灰石复性及同时纯化基因重组蛋白质的方法 |
CN101153050A (zh) * | 2006-09-29 | 2008-04-02 | 中国科学院海洋研究所 | 一种分离纯化组蛋白h4的方法 |
WO2008127305A2 (en) | 2006-11-01 | 2008-10-23 | Biogen Idec Ma Inc. | Method of isolating biomacromolecules using low ph and divalent cations |
US8163481B2 (en) * | 2007-04-02 | 2012-04-24 | University of Pittsburgh - of the Commonwealth Systems of Higher Education | Method of extracting chromatin fractions from intact cells |
US8329871B2 (en) | 2008-06-24 | 2012-12-11 | Octapharma Ag | Process of purifying coagulation factor VIII |
RU2408602C1 (ru) * | 2009-04-22 | 2011-01-10 | Учреждение Российской академии наук Институт биологии Уфимского научного центра РАН (ИБ УНЦ РАН) | Способ препаративного выделения основных белков из супраструктур клеточных ядер растений |
EP2855504B1 (en) * | 2012-05-31 | 2018-10-24 | Agency For Science, Technology And Research | Chromatographic purification of immunoglobulin g preparations with particles having multimodal functionalities |
WO2013180648A1 (en) * | 2012-05-31 | 2013-12-05 | Agency For Science, Technology And Research | Methods for use of mixed multifunctional surfaces for reducing aggregate content in protein preparations |
-
2014
- 2014-05-29 EP EP14893573.7A patent/EP3149023A4/en not_active Withdrawn
- 2014-05-29 CN CN201480078439.4A patent/CN106232614A/zh active Pending
- 2014-05-29 SG SG11201609815QA patent/SG11201609815QA/en unknown
- 2014-05-29 US US15/313,329 patent/US10259851B2/en not_active Expired - Fee Related
- 2014-05-29 JP JP2017515662A patent/JP6599980B2/ja not_active Expired - Fee Related
- 2014-05-29 WO PCT/SG2014/000237 patent/WO2015183183A1/en active Application Filing
- 2014-05-29 KR KR1020167028474A patent/KR20170010750A/ko not_active Application Discontinuation
Also Published As
Publication number | Publication date |
---|---|
CN106232614A (zh) | 2016-12-14 |
EP3149023A1 (en) | 2017-04-05 |
JP2017519044A (ja) | 2017-07-13 |
EP3149023A4 (en) | 2017-11-22 |
US10259851B2 (en) | 2019-04-16 |
US20170137479A1 (en) | 2017-05-18 |
SG11201609815QA (en) | 2016-12-29 |
KR20170010750A (ko) | 2017-02-01 |
WO2015183183A1 (en) | 2015-12-03 |
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