{"title":"Refolding Proteins from Inclusion Bodies using Differential Scanning Fluorimetry Guided (DGR) Protein Refolding and MeltTraceur Web","authors":"Mark E. Lee, Xiaoyi Dou, Yingmin Zhu, Kevin J. Phillips","doi":"10.1002/cpmb.78","DOIUrl":null,"url":null,"abstract":"<p>Differential Scanning Fluorimetry Guided Refolding (DGR) is a simple methodology that can be used to rapidly screen for and identify conditions capable of accurately refolding protein preparations, such as those obtained from <i>Escherichia coli</i> inclusion bodies. It allows for the production in <i>E. coli</i> of functional proteins that would otherwise require far more expensive production methods. This unit describes how to set up a DGR refolding assay, perform DGR refolding trials in microplate format, use MeltTraceur Web software to interactively analyze the resulting data, scale-up protein production via refolding, and lastly, validate that the protein is properly folded. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":10734,"journal":{"name":"Current Protocols in Molecular Biology","volume":"125 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpmb.78","citationCount":"6","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Molecular Biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpmb.78","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 6
Abstract
Differential Scanning Fluorimetry Guided Refolding (DGR) is a simple methodology that can be used to rapidly screen for and identify conditions capable of accurately refolding protein preparations, such as those obtained from Escherichia coli inclusion bodies. It allows for the production in E. coli of functional proteins that would otherwise require far more expensive production methods. This unit describes how to set up a DGR refolding assay, perform DGR refolding trials in microplate format, use MeltTraceur Web software to interactively analyze the resulting data, scale-up protein production via refolding, and lastly, validate that the protein is properly folded. © 2018 by John Wiley & Sons, Inc.
利用差示扫描荧光法引导(DGR)蛋白质重折叠和熔融示踪网从包涵体中重折叠蛋白质
差示扫描荧光法引导重折叠(DGR)是一种简单的方法,可用于快速筛选和确定能够准确重折叠蛋白质制剂的条件,例如从大肠杆菌包涵体中获得的蛋白质制剂。它允许在大肠杆菌中生产功能性蛋白质,否则需要更昂贵的生产方法。本单元描述了如何建立DGR重折叠实验,在微孔板格式中进行DGR重折叠试验,使用MeltTraceur Web软件交互分析结果数据,通过重折叠扩大蛋白质产量,最后验证蛋白质是否正确折叠。©2018 by John Wiley &儿子,Inc。
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