• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利:CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法,用中文像聊天一样搜索,搜遍4000万医学文献。AI智能推荐,让科研检索更轻松。

立即免费搜索

文件翻译

保留排版,准确专业,支持PDF/Word/PPT等文件格式,支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述,25分钟生成高质量综述,智能提取关键信息,辅助科研写作。

立即免费体验

用有机溶剂溶解人干扰素β-1b包涵体蛋白

Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents.

作者信息

Nekoufar Samira, Fazeli Ahmad, Fazeli Mohammad Reza

机构信息

Department of Biochemistry, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.

The Institute of Pharmaceutical Sciences (TIPS), Tehran University of Medical Sciences (TUMS), Tehran, Iran.

出版信息

Adv Pharm Bull. 2020 Jun;10(2):233-238. doi: 10.34172/apb.2020.027. Epub 2020 Feb 18.

DOI:10.34172/apb.2020.027
PMID:32373491
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7191233/
Abstract

Solubilization of inclusion bodies expressed in is a critical step during manufacturing of recombinant proteins expressed as inclusion bodies. So far, various methods have been used for solubilization and purification of inclusion body proteins to obtain active proteins with high purity and yield. The aim of this study was to examine the benefit of organic solvents such as alcohols in solubilization of recombinant interferon β-1b inclusion bodies. Effect of important parameters inclusion pH, concentration and type of denaturant and concentration of alcoholic solvents were optimized to formulate a suitable solubilization buffer and investigate their effect on solubilization of interferon β-1b inclusion bodies. Our findings showed the acidic pH in the range of 2-3 is more suitable than alkaline pH >12 for solubilization and achieving higher content of interferon β-1beta and pure recombinant protein. We have also demonstrated that 1% SDS acts better than 2M urea to solubilize Inclusion body proteins of interferon β-1b at pH of 2-3. The interferon concentration was 2.35 mg per 100 mg IB when we used 40% () 1-propanol and 20% () 2-butanol into the buffer solution as well. The optimized method provides gentile condition for solubilization of inclusion body at high protein concentration and purity with a degree of retention of native secondary structure which makes this method valuable to be used in production and research area.

摘要

在重组蛋白以包涵体形式表达的生产过程中,包涵体的溶解是关键步骤。到目前为止,已采用多种方法来溶解和纯化包涵体蛋白,以获得高纯度和高产量的活性蛋白。本研究旨在探讨醇类等有机溶剂在重组干扰素β-1b包涵体溶解中的作用。优化了包括pH值、变性剂浓度和类型以及醇类溶剂浓度等重要参数的影响,以配制合适的溶解缓冲液,并研究它们对干扰素β-1b包涵体溶解的影响。我们的研究结果表明,pH值在2 - 3范围内的酸性条件比pH值>12的碱性条件更适合溶解,并且能获得更高含量的干扰素β-1β和纯重组蛋白。我们还证明,在pH值为2 - 3时,1% SDS比2M尿素在溶解干扰素β-1b包涵体蛋白方面效果更好。当我们在缓冲溶液中使用40%(体积分数)1-丙醇和20%(体积分数)2-丁醇时,每100 mg包涵体中的干扰素浓度为2.35 mg。该优化方法为在高蛋白浓度和纯度下溶解包涵体提供了温和的条件,并保留了一定程度的天然二级结构,这使得该方法在生产和研究领域具有应用价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/d2a8d48db535/apb-10-233-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/103e18aead4f/apb-10-233-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/c04ff5e0b735/apb-10-233-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/cfebccfc87f4/apb-10-233-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/cabdac85deda/apb-10-233-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/d2a8d48db535/apb-10-233-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/103e18aead4f/apb-10-233-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/c04ff5e0b735/apb-10-233-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/cfebccfc87f4/apb-10-233-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/cabdac85deda/apb-10-233-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2d4e/7191233/d2a8d48db535/apb-10-233-g006.jpg

相似文献

1
Solubilization of Human Interferon β-1b Inclusion Body Proteins by Organic Solvents.用有机溶剂溶解人干扰素β-1b包涵体蛋白
Adv Pharm Bull. 2020 Jun;10(2):233-238. doi: 10.34172/apb.2020.027. Epub 2020 Feb 18.
2
Recovery of bioactive protein from bacterial inclusion bodies using trifluoroethanol as solubilization agent.以三氟乙醇为增溶剂从细菌包涵体中回收生物活性蛋白。
Microb Cell Fact. 2016 Jun 8;15:100. doi: 10.1186/s12934-016-0504-9.
3
[Solubilization Specificities Interferon beta-1b from Inclusion Bodies].[包涵体中β-1b干扰素的溶解特性]
Bioorg Khim. 2015 Jul-Aug;41(4):403-10. doi: 10.1134/s1068162015040159.
4
A single freeze-thawing cycle for highly efficient solubilization of inclusion body proteins and its refolding into bioactive form.单次冻融循环可高效溶解包涵体蛋白并将其重折叠为生物活性形式。
Microb Cell Fact. 2015 Feb 22;14:24. doi: 10.1186/s12934-015-0208-6.
5
The use of Zwittergent 3-14 in the purification of recombinant human interferon-beta Ser17 (Betaseron).两性离子去污剂3-14在重组人干扰素-β Ser17(倍泰龙)纯化中的应用
J Interferon Cytokine Res. 1995 Jan;15(1):31-7. doi: 10.1089/jir.1995.15.31.
6
High pH solubilization and chromatography-based renaturation and purification of recombinant human granulocyte colony-stimulating factor from inclusion bodies.高 pH 溶解结合基于色谱的复性和从包涵体中纯化重组人粒细胞集落刺激因子。
Appl Biochem Biotechnol. 2012 Mar;166(5):1264-74. doi: 10.1007/s12010-011-9512-8. Epub 2012 Jan 3.
7
Capillary gel electrophoresis for the quantification and purity determination of recombinant proteins in inclusion bodies.毛细管凝胶电泳法用于测定包涵体中重组蛋白的定量和纯度。
Electrophoresis. 2013 Sep;34(18):2754-9. doi: 10.1002/elps.201300232. Epub 2013 Aug 13.
8
Effect of buffer additives on solubilization and refolding of reteplase inclusion bodies.缓冲液添加剂对瑞替普酶包涵体溶解和复性的影响。
Res Pharm Sci. 2018 Oct;13(5):413-421. doi: 10.4103/1735-5362.236834.
9
Effect of post-solubilization conditions on the yield and efficiency of recombinant streptokinase purification at large-scale.后增溶条件对大规模重组链激酶纯化产量和效率的影响。
Protein Expr Purif. 2014 Sep;101:127-32. doi: 10.1016/j.pep.2014.06.007. Epub 2014 Jun 17.
10
Association of high pressure and alkaline condition for solubilization of inclusion bodies and refolding of the NS1 protein from zika virus.高压和碱性条件对 Zika 病毒 NS1 蛋白包涵体溶解和复性的影响。
BMC Biotechnol. 2018 Dec 12;18(1):78. doi: 10.1186/s12896-018-0486-2.

引用本文的文献

1
State-of-the-art and novel approaches to mild solubilization of inclusion bodies.包涵体温和溶解的先进与新颖方法。
Front Bioeng Biotechnol. 2023 Jul 20;11:1249196. doi: 10.3389/fbioe.2023.1249196. eCollection 2023.
2
Strategic optimization of conditions for the solubilization of GST-tagged amphipathic helix-containing ciliary proteins overexpressed as inclusion bodies in E. coli.原核表达 GST 标签的两亲性螺旋亲水融合蛋白包涵体的增溶条件的优化策略。
Microb Cell Fact. 2022 Dec 12;21(1):258. doi: 10.1186/s12934-022-01979-y.
3
Interferon-Based Biopharmaceuticals: Overview on the Production, Purification, and Formulation.

本文引用的文献

1
Recovery of bioactive protein from bacterial inclusion bodies using trifluoroethanol as solubilization agent.以三氟乙醇为增溶剂从细菌包涵体中回收生物活性蛋白。
Microb Cell Fact. 2016 Jun 8;15:100. doi: 10.1186/s12934-016-0504-9.
2
[Solubilization Specificities Interferon beta-1b from Inclusion Bodies].[包涵体中β-1b干扰素的溶解特性]
Bioorg Khim. 2015 Jul-Aug;41(4):403-10. doi: 10.1134/s1068162015040159.
3
Pillars Article: Virus Interference. I. The Interferon. Proc R Soc Lond B Biol Sci. 1957. 147: 258-267.支柱文章:病毒干扰。I. 干扰素。《伦敦皇家学会学报B辑:生物科学》。1957年。第147卷:第258 - 267页。
基于干扰素的生物制药:生产、纯化及制剂概述
Vaccines (Basel). 2021 Apr 1;9(4):328. doi: 10.3390/vaccines9040328.
J Immunol. 2015 Sep 1;195(5):1911-20.
4
Protein recovery from inclusion bodies of Escherichia coli using mild solubilization process.采用温和溶解法从大肠杆菌包涵体中回收蛋白质。
Microb Cell Fact. 2015 Mar 25;14:41. doi: 10.1186/s12934-015-0222-8.
5
Recombinant protein expression in Escherichia coli: advances and challenges.大肠杆菌中的重组蛋白表达:进展与挑战
Front Microbiol. 2014 Apr 17;5:172. doi: 10.3389/fmicb.2014.00172. eCollection 2014.
6
The mechanism of action of interferon-β in relapsing multiple sclerosis.干扰素-β在复发型多发性硬化症中的作用机制。
CNS Drugs. 2011 Jun 1;25(6):491-502. doi: 10.2165/11591110-000000000-00000.
7
Inclusion bodies: a new concept.包涵体:新概念。
Microb Cell Fact. 2010 Nov 1;9:80. doi: 10.1186/1475-2859-9-80.
8
Cloning, high expression and purification of recombinant human intereferon-beta-1b in Escherichia coli.重组人干扰素-β-1b在大肠杆菌中的克隆、高效表达及纯化
Appl Biochem Biotechnol. 2009 Jul;158(1):140-54. doi: 10.1007/s12010-008-8318-9. Epub 2008 Aug 5.
9
Isolation, renaturation, and formation of disulfide bonds of eukaryotic proteins expressed in Escherichia coli as inclusion bodies.以包涵体形式在大肠杆菌中表达的真核蛋白质的分离、复性及二硫键形成
Biotechnol Bioeng. 1993 Jan 5;41(1):3-13. doi: 10.1002/bit.260410103.
10
Production of active eukaryotic proteins through bacterial expression systems: a review of the existing biotechnology strategies.通过细菌表达系统生产活性真核蛋白:现有生物技术策略综述
Mol Cell Biochem. 2008 Jan;307(1-2):249-64. doi: 10.1007/s11010-007-9603-6. Epub 2007 Sep 12.