JP6771750B2 - 生体分子の回収、安定化及び溶出のための基材及び方法 - Google Patents
生体分子の回収、安定化及び溶出のための基材及び方法 Download PDFInfo
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- JP6771750B2 JP6771750B2 JP2016563056A JP2016563056A JP6771750B2 JP 6771750 B2 JP6771750 B2 JP 6771750B2 JP 2016563056 A JP2016563056 A JP 2016563056A JP 2016563056 A JP2016563056 A JP 2016563056A JP 6771750 B2 JP6771750 B2 JP 6771750B2
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
-
- 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
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
- C07K1/113—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure
- C07K1/1136—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure by reversible modification of the secondary, tertiary or quarternary structure, e.g. using denaturating or stabilising agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K17/00—Carrier-bound or immobilised peptides; Preparation thereof
- C07K17/02—Peptides being immobilised on, or in, an organic carrier
- C07K17/10—Peptides being immobilised on, or in, an organic carrier the carrier being a carbohydrate
- C07K17/12—Cellulose or derivatives thereof
-
- 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
- C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
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- Genetics & Genomics (AREA)
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- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
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- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
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- Bioinformatics & Cheminformatics (AREA)
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- General Engineering & Computer Science (AREA)
- Biotechnology (AREA)
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- General Physics & Mathematics (AREA)
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- Crystallography & Structural Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Peptides Or Proteins (AREA)
- Sampling And Sample Adjustment (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Description
試薬:31−ETFはGE Healthcareから入手した。セルロース紙(Whatman 31ETF)を、適切な配合物の暖めた溶液に浸漬し、その後ラインオーブンコンベヤを使用してこの基材を乾燥させることによって、紙基材にメレジトース及び他の試薬を含浸させた。乾燥基材をその後、防湿剤と共に後の試験までMylarバッグ中に密閉した。4種の浸漬用配合物を調製した:(1)15%(重量/体積基準)メレジトース溶液、(2)15%メレジトースも含有する標準FTA溶液。標準FTA成分は、下記(重量/体積基準):0.24%EDTA、1.63%ドデシル硫酸ナトリウム(SDS)、1.61%Tris緩衝液塩及び0.56%尿酸、(3)5%(重量/体積基準)Ficoll PM400及び(4)下記のパーセントの小成分を含有した溶液:6.5%メレジトース、4.2%Ficoll PM70及び4.2%Ficoll PM400を含み、これ以降メレジトース−Ficoll配合物と称される。
3種のタンパク質を、セルロース系基材上の初期安定性評価のために選択した:サイトカインIL−8、コレステロールタンパク質アポリポタンパク質B(ApoB)及び酵素β−ガラクトシダーゼ(β−gal)。IL−8は、呼吸器感染症の代表的バイオマーカーであるため選択した。ApoBは、易動性タンパク質モデル(すなわち、乾燥血液スポットにおいて半減期が短いことが公知である)として選択した。β−galは、酵素活性の直接定量を提供するその能力のため、第3のモデルタンパク質として選択した。
配合物のリストは、長期(90日)試料貯蔵後のタンパク質の安定性を決定するために選択した。本実施例では、目的の分析物を含有するヒト血液試料を各基材に適用し、低湿度条件下で30℃において90日間貯蔵し、その後実施例2に記載のようにタンパク質の安定性を決定した。全基材を実施例1に上記のように調製した。基材配合物のパンチに、血液中のIL−1β(6pg/μL)、IL−8(7.5ng/mL)、TNF−α(8pg/μL)又はβ−gal(10μg/mL)を個別に添加し、上記のように室温で乾燥させた。選択された基材由来の乾燥血液スポットを、30℃における長期貯蔵(丸括弧内は貯蔵日数)後、未修飾31−ETFセルロースと比較した。
サイトカイン標的(IL−1β、IL−8及びTNF−α)及びApoBに関するELISAシグナルは、様々なセルロース基材の相対的安定化効果の有用な測定を提供した。分析物のシグナルが抑制される場合、実質的なタンパク質の変性と相まった完全な溶出からもたらされるのか、又は実質的に活性な非変性タンパク質の不完全な溶出からもたらされるのかどうかを決定することは困難である。β−galシグナルは必然的に活性タンパク質に依存するので、この特定の分析物は、シグナルの「理論値」とシグナルの溶出による実測値とを比較することによって、活性な非変性タンパク質の実際の溶出効率を測定する機会を提供した。
Claims (6)
- タンパク質、酵素、サイトカイン又はこれらの組合せの安定化及び固体基材からのタンパク質、酵素、サイトカイン又はこれらの組合せの溶出のための、メレジトースで含浸された固体基材であって、
前記基材がセルロース膜であり、
メレジトースの濃度が15%である、
基材。 - 1種以上の溶解剤をさらに含み、溶解剤が洗浄剤、カオトロープ、変性剤又はこれらの組合せを含み、チオシアン酸塩、陰イオン性洗浄剤、非イオン性洗浄剤、陽イオン性洗浄剤、尿素又はこれらの組合せから選択される、請求項1に記載の基材。
- 還元剤、緩衝剤、抗酸化剤、キレート剤又はこれらの組合せをさらに含み、還元剤がジチオスレイトール(DTT)、2−メルカプトエタノール(2−ME)、トリス(2−カルボキシエチル)ホスフィン(TCEP)又はこれらの組合せから選択され、緩衝剤が2−アミノ−2−ヒドロキシメチル−プロパン−1,3−ジオール(Tris)、2−(N−モルホリノ)エタンスルホン酸(MES)、3−(N−モルホリノ)プロパンスルホン酸(MOPS)、クエン酸緩衝剤、4−(2−ヒドロキシエチル)−1−ピペラジンエタンスルホン酸(HEPES)、リン酸緩衝剤又はこれらの組合せから選択され、キレート剤が、エチレンジアミン四酢酸(EDTA)、クエン酸、エチレングリコール四酢酸(EGTA)又はこれらの組合せから選択される、請求項1又は請求項2に記載の基材。
- デキストラン、フィコール、キトサン、アミロペクチン、アルギン酸、カルボキシメチルセルロース又はこれらの組合せから選択される多糖類をさらに含む請求項1乃至請求項3のいずれか1項に記載の基材。
- 周囲温度でタンパク質、酵素、サイトカイン又はこれらの組合せの安定性及び溶出率を向上さるように構成される、請求項1乃至請求項4のいずれか1項に記載の基材。
- 20〜22℃の周囲温度で、少なくとも1乃至3ヶ月の貯蔵期間におけるタンパク質、酵素、サイトカイン又はこれらの組合せの安定性を向上させるように構成される、請求項1乃至請求項5のいずれか1項に記載の基材。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US14/261,900 US9480966B2 (en) | 2012-04-30 | 2014-04-25 | Substrates and methods for collection, stabilization and elution of biomolecules |
US14/261,900 | 2014-04-25 | ||
PCT/EP2015/058518 WO2015162093A1 (en) | 2014-04-25 | 2015-04-20 | Substrates and methods for collection, stabilization and elution of biomolecules |
Publications (2)
Publication Number | Publication Date |
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JP2017522263A JP2017522263A (ja) | 2017-08-10 |
JP6771750B2 true JP6771750B2 (ja) | 2020-10-21 |
Family
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Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2016563056A Active JP6771750B2 (ja) | 2014-04-25 | 2015-04-20 | 生体分子の回収、安定化及び溶出のための基材及び方法 |
Country Status (11)
Country | Link |
---|---|
EP (1) | EP3134441B1 (ja) |
JP (1) | JP6771750B2 (ja) |
KR (1) | KR102495546B1 (ja) |
CN (2) | CN115144512A (ja) |
AU (2) | AU2015250915B2 (ja) |
CA (1) | CA2944876A1 (ja) |
HK (1) | HK1232291A1 (ja) |
IL (1) | IL248052B (ja) |
MX (1) | MX2016013980A (ja) |
SG (1) | SG11201608021TA (ja) |
WO (1) | WO2015162093A1 (ja) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
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US9480966B2 (en) | 2012-04-30 | 2016-11-01 | General Electric Company | Substrates and methods for collection, stabilization and elution of biomolecules |
EP4035762B1 (en) * | 2015-09-09 | 2023-11-01 | Drawbridge Health, Inc. | Devices for sample collection, stabilization and preservation |
SG10201911882VA (en) | 2017-01-10 | 2020-02-27 | Drawbridge Health Inc | Devices, systems, and methods for sample collection |
CN108579671B (zh) * | 2018-01-23 | 2020-08-25 | 辽宁大学 | 一种用于重金属离子吸附的碳量子点及其制备方法和应用 |
CN110878115B (zh) * | 2019-12-11 | 2021-08-03 | 湖南师范大学 | 蛋白质提取液、等电聚焦裂解液及其在蛋白质提取、等电聚焦及非变性凝胶电泳中的应用 |
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JP2728757B2 (ja) * | 1988-02-11 | 1998-03-18 | キューノ,インコーポレーテッド | 変性多糖支持体の親和性マトリックス |
GB9010742D0 (en) * | 1990-05-14 | 1990-07-04 | Quadrant Bioresources Ltd | Stabilization of biological macromolecular substances |
WO2003020924A2 (en) * | 2001-09-05 | 2003-03-13 | Whatman Plc | Stable storage of proteins |
WO2003087335A2 (en) * | 2002-04-11 | 2003-10-23 | Medimmune Vaccines, Inc. | Preservation of bioactive materials by spray drying |
WO2003086443A1 (en) * | 2002-04-11 | 2003-10-23 | Medimmune Vaccines, Inc. | Spray freeze dry of compositions for intranasal administration |
DE10225501A1 (de) * | 2002-06-10 | 2003-12-24 | Eppendorf Ag | Verfahren zur Immobilisierung von in einer flüssigen Probe enthaltenen Proteinen mit anschließender Trocknung der immobilisierten Proteine, sowie in dem Verfahren einsetzbare Festphasensubstrate |
US20080176209A1 (en) * | 2004-04-08 | 2008-07-24 | Biomatrica, Inc. | Integration of sample storage and sample management for life science |
CA2761675A1 (en) * | 2009-05-11 | 2010-11-18 | Biomatrica, Inc. | Compositions and methods for biological sample storage |
WO2010144682A1 (en) * | 2009-06-12 | 2010-12-16 | Micronics, Inc. | Rehydratable matrices for dry storage of taq polymerase in a microfluidic device |
CN102947082B (zh) * | 2010-04-06 | 2016-03-30 | 金沃特公司 | 生物材料的稳定化的化学脱水 |
KR101872203B1 (ko) * | 2010-04-20 | 2018-08-02 | 옥타파마 아게 | 약학 단백질을 위한 신규한 안정화제 |
GB201103256D0 (en) * | 2011-02-25 | 2011-04-13 | Ge Healthcare Uk Ltd | Solid support and method of recovering biological material therefrom |
AU2013249007B2 (en) * | 2012-04-20 | 2016-04-21 | California Institute Of Technology | Fluidic devices and systems for sample preparation or autonomous analysis |
CN102728407B (zh) * | 2012-07-17 | 2014-06-11 | 岳阳亚王精细化工有限公司 | 一种(S,S)-salen Co(Ⅱ)催化剂的合成方法及其在拆分末端环氧化合物的应用 |
US20140038172A1 (en) * | 2012-08-06 | 2014-02-06 | Vivebio, Llc | Matrix and System for Preserving Biological Specimens for Qualitative and Quantitative Analysis |
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2015
- 2015-04-20 KR KR1020167029315A patent/KR102495546B1/ko active IP Right Grant
- 2015-04-20 WO PCT/EP2015/058518 patent/WO2015162093A1/en active Application Filing
- 2015-04-20 EP EP15717489.7A patent/EP3134441B1/en active Active
- 2015-04-20 MX MX2016013980A patent/MX2016013980A/es active IP Right Grant
- 2015-04-20 CA CA2944876A patent/CA2944876A1/en active Pending
- 2015-04-20 CN CN202210786242.2A patent/CN115144512A/zh active Pending
- 2015-04-20 SG SG11201608021TA patent/SG11201608021TA/en unknown
- 2015-04-20 AU AU2015250915A patent/AU2015250915B2/en active Active
- 2015-04-20 CN CN201580021334.XA patent/CN106233137A/zh active Pending
- 2015-04-20 JP JP2016563056A patent/JP6771750B2/ja active Active
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2016
- 2016-09-26 IL IL248052A patent/IL248052B/en unknown
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2017
- 2017-06-14 HK HK17105875.0A patent/HK1232291A1/zh unknown
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Also Published As
Publication number | Publication date |
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AU2015250915B2 (en) | 2018-02-08 |
WO2015162093A1 (en) | 2015-10-29 |
HK1232291A1 (zh) | 2018-01-05 |
CN106233137A (zh) | 2016-12-14 |
CN115144512A (zh) | 2022-10-04 |
AU2015250915A1 (en) | 2016-10-20 |
IL248052B (en) | 2021-12-01 |
MX2016013980A (es) | 2017-01-11 |
KR102495546B1 (ko) | 2023-02-06 |
IL248052A0 (en) | 2016-11-30 |
EP3134441A1 (en) | 2017-03-01 |
AU2018203078A1 (en) | 2018-05-24 |
EP3134441B1 (en) | 2019-12-18 |
CA2944876A1 (en) | 2015-10-29 |
SG11201608021TA (en) | 2016-11-29 |
KR20160145035A (ko) | 2016-12-19 |
JP2017522263A (ja) | 2017-08-10 |
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