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{"title":"Optimization of Protein Expression in Mammalian Cells","authors":"Molly Hunter, Ping Yuan, Divya Vavilala, Mark Fox","doi":"10.1002/cpps.77","DOIUrl":null,"url":null,"abstract":"<p>Recombinant proteins, such as monoclonal antibodies, are produced in mammalian cell lines to introduce proper protein folding and post-translational modifications, which are essential for full biological activity. In both the industrial and academic environments, the use of recombinant proteins varies widely and, with it, the method of production. The amount of an antibody needed for a toxicity study is far greater than that needed by a research lab performing cellular assays, and the amount of effort put into the development of the protein will vary accordingly. There is no universal strategy for mammalian expression systems, and scientists often struggle to develop a suitable process from the myriad of choices at each step. Here, we elaborate on the various obstacles encountered when planning high-yield experiments to produce the recombinant proteins of interest. © 2018 by John Wiley & Sons, Inc.</p>","PeriodicalId":10866,"journal":{"name":"Current Protocols in Protein Science","volume":"95 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpps.77","citationCount":"91","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Protein Science","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpps.77","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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Abstract
Recombinant proteins, such as monoclonal antibodies, are produced in mammalian cell lines to introduce proper protein folding and post-translational modifications, which are essential for full biological activity. In both the industrial and academic environments, the use of recombinant proteins varies widely and, with it, the method of production. The amount of an antibody needed for a toxicity study is far greater than that needed by a research lab performing cellular assays, and the amount of effort put into the development of the protein will vary accordingly. There is no universal strategy for mammalian expression systems, and scientists often struggle to develop a suitable process from the myriad of choices at each step. Here, we elaborate on the various obstacles encountered when planning high-yield experiments to produce the recombinant proteins of interest. © 2018 by John Wiley & Sons, Inc.
哺乳动物细胞中蛋白表达的优化
重组蛋白,如单克隆抗体,在哺乳动物细胞系中产生,以引入适当的蛋白质折叠和翻译后修饰,这是充分的生物活性所必需的。在工业和学术环境中,重组蛋白的使用变化很大,生产方法也随之变化。毒性研究所需的抗体量远远大于进行细胞分析的研究实验室所需的抗体量,因此用于开发蛋白质的努力量也会相应变化。哺乳动物的表达系统并没有通用的策略,科学家们经常努力从每一步的无数选择中找到一个合适的过程。在这里,我们详细说明了在计划生产感兴趣的重组蛋白的高产实验时遇到的各种障碍。©2018 by John Wiley &儿子,Inc。
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