Use of MAR Elements to Increase the Production of Recombinant Proteins
Use of MAR Elements to Increase the Production of Recombinant Proteins
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DOI：
10.1007/978-90-481-2245-5_1
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发表时间：
2009-07-19
期刊：
影响因子：
--
通讯作者：
Mermod N
中科院分区：
文献类型：
--
作者：
Gorman C;Arope S;Grandjean M;Girod PA;Mermod N
生物制药行业继续面临生产大量用作治疗药物的重组蛋白的挑战，临床开发中百分之八十的蛋白质治疗药物是在哺乳动物细胞系统中生产的。在过去的几十年中，针对生长条件提高产量的方法，例如改进培养基组成和过程控制，或通过使用强启动子/增强子和扩增基因拷贝数来转录重组基因，已经大大提高了从哺乳动物细胞获得的产量。然而，这些过程仍然很费力，并且通常需要对克隆进行广泛的筛选，因为并不总能获得稳定的细胞系和/或蛋白质生产。不稳定或可变的表达与转基因整合位点的位置、基因表达的调节、基因的沉默和基因拷贝的丢失有关。可以重塑染色质以维持转基因处于活性构型的遗传元件现在越来越多地被用来提高哺乳动物细胞的蛋白质产量。在这里，我们将回顾其中一种类型的元件（MAR）如何增加转基因整合到细胞基因组中并减少沉默效应以减少表达变异性。我们还说明了在表达载体中包含这些元件如何导致每个细胞每天 (p/c/d) 提高 20 至 100 皮克的比生产率，从而使蛋白质滴度高于 5 g/l。
The biopharmaceutical industry continues to face the challenge of producing large amount of recombinant proteins for use as therapeutics, and eighty percent of protein therapeutics in clinical development are produced in mammalian cell systems. Approaches to increase production addressing growth conditions, such as the improvement of media composition and process control, or transcription of the recombinant gene via the use of strong promoters/enhancers and amplification of gene copy number, have increased the yields obtained from mammalian cells considerably over the past decades. However these processes remain laborious, and extensive screening of clones is often required, as stable cell line and/or protein production is not always obtained. Unstable or variable expression is linked to the location of transgene integration site, the regulation of gene expression, the silencing of genes, and the loss of gene copies. Genetic elements that may remodel chromatin to maintain the transgene in an active configuration are now being employed increasingly to improve protein production using mammalian cells. Here we will review how one type of such elements, the MARs, may increase transgene integration into the cell genome and decrease silencing effects to reduce expression variability. We also illustrate how inclusion of these elements in expression vectors leads to increased specific productivities ranging from 20 to 100 picograms per cell and per day (p/c/d), resulting in protein titers above 5 g/l.
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