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The use of peptones as medium additives for the production of a recombinant therapeutic protein in high density perfusion cultures of mammalian cells

机译:蛋白ept作为培养基添加剂在哺乳动物细胞高密度灌注培养中生产重组治疗蛋白的用途

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摘要

Protein hydrolysates as substitutes for serum havebeen employed by many in cell culture mediumformulation, especially with the shift to low proteinor protein-free media. More recently, vegetablehydrolysates have also been added as nutritionalsupplements to fortify the amino acid content in smallpeptide form for batch and fed-batch fermentations. Several of these new hydrolysates (peptones of soy,rice, wheat gluten etc.) were tested as protein-freemedium supplements for the production of a recombinanttherapeutic protein. Multiple peptone-supplemented,continuous perfusion bioreactor experiments wereconducted, varying dilution rates and basal mediumcomposition over the various runs. Cell specificrates and product quality studies were obtained forthe various peptones and compared with peptone-freemedium. The potential for peptones to decreaseintrinsic and proteolytic degradation of the productwas also investigated.It was found that peptones confer a nutritionalbenefit, especially at low dilution rates, for therecombinant BHK cell line used in this investigation.The specific productivity increased 20–30% comparedto the peptone-free controls. However, this benefitwas also fully delivered by using fortified medium inplace of the peptone-enriched media. Therefore, whilepeptones may be considered as useful medium additiveswhen development time is limited, their addition maybe avoided by systematic medium development ifpermitted by the time line of the project.
机译:在细胞培养基的配制中,许多人已经采用蛋白质水解物代替血清,尤其是向低蛋白质或无蛋白质的培养基转移。最近,还添加了植物水解物作为营养补充品,以增强小肽形式的氨基酸含量,用于分批和补料分批发酵。已测试了其中几种新的水解产物(大豆,大米,小麦面筋等蛋白)作为无蛋白培养基的补充品,用于生产重组治疗性蛋白质。进行了多个补充蛋白up的连续灌注生物反应器实验,在不同的运行过程中改变了稀释率和基础培养基组成。获得了各种蛋白ept的细胞特异性和产品质量研究,并与无蛋白med的培养基进行了比较。还研究了蛋白to降低产品内在和蛋白水解降解的潜力。发现,蛋白investigation为这项研究中使用的BHK重组细胞系提供了营养益处,尤其是在低稀释度时。比生产率提高了20–30%。无蛋白controls的对照。但是,通过使用强化培养基代替富含蛋白ept的培养基,也可以完全实现这种益处。因此,在开发时间有限的情况下,pe可能被视为有用的培养基添加剂,但如果项目时间允许,可以通过系统的培养基开发来避免添加pe。

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