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首页> 外文期刊>Biotechnology Progress >Proteomic Analysis for the Assessment of Different Lots of Fetal Bovine Serum as a Raw Material for Cell Culture.Part IV.Application of Proteomics to the Manufacture of Biological Drugs
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Proteomic Analysis for the Assessment of Different Lots of Fetal Bovine Serum as a Raw Material for Cell Culture.Part IV.Application of Proteomics to the Manufacture of Biological Drugs

机译：蛋白质组学分析对不同批次胎牛血清作为细胞培养原料的评估。第四部分蛋白质组学在生物药物生产中的应用

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Fetal bovine serum (FBS) is the most widely used growth supplement for cell cultures,primarily because of its high levels of growth stimulatory factors and low levels of growth inhibitory factors.Maintaining successful and consistent cell fermentations can be difficult,as FBS is a complex natural product and may vary from lot to lot even from a single manufacturer.The quality and concentration of both bulk and specific proteins can affect cell growth.Quality control tools for FBS are relatively primitive and expensive given the complexity of the sample and the large amounts of FBS used.We undertook this study to examine whether proteomics could be used as a tool to analyze the variability of different fermentation processes.We hypothesized that inconsistent cell growth in fermentations could be due to the quality of FBS and that different lots of FBS had varying concentrations of proteins such as growth stimulatory factors,growth inhibitory factors,and/or other proteins that may correlate with cellular growth rate.To investigate whether this was the case,we grew three batches of adult retinal pigment epithelial cells (ARPE-19) using three different lots of fetal bovine serum (FBS-Ia,FBS-Ib,and FBS-II).We found that the growth rate of the culture was significantly and consistently higher in the FBS-II lot.To determine why the other lots promoted different growth properties,we used proteomic techniques to analyze the protein composition of the three lots.We then performed a time course study to monitor specific changes in individual proteins in the fermentation medium.The amount of several extracellular matrix and structural proteins,which are indicators of cell growth,increased over time.Alternatively,components supplied by the FBS addition,such as nutritional-related and cell-spreading-related proteins,decreased over time.
机译：胎牛血清（FBS）是细胞培养中使用最广泛的生长补品，主要是因为它具有高水平的生长刺激因子和低水平的生长抑制因子。由于FBS复杂，难以维持成功且持续的细胞发酵天然产物，甚至同一家制造商的批次之间也可能会有所不同。大块蛋白质和特定蛋白质的质量和浓度都会影响细胞生长。鉴于样品的复杂性和数量庞大，FBS的质量控制工具相对原始且昂贵我们进行了这项研究，以研究蛋白质组学是否可以用作分析不同发酵过程变异性的工具。我们假设发酵中细胞生长不一致可能是由于FBS的质量造成的，并且不同批次的FBS具有不同浓度的蛋白质，例如生长刺激因子，生长抑制因子和/或其他可能相关的蛋白质为了研究是否如此，我们使用三批不同的胎牛血清（FBS-Ia，FBS-Ib和FBS-II）培养了三批成年视网膜色素上皮细胞（ARPE-19）。 ）。我们发现FBS-II批次的培养物生长速度显着且始终较高。为确定其他批次为何促进了不同的生长特性，我们使用了蛋白质组学技术来分析这三个批次的蛋白质组成。进行了一个时程研究，以监测发酵培养基中各个蛋白质的特定变化。几种指示细胞生长的细胞外基质和结构蛋白的量随时间的推移而增加。相关蛋白和细胞扩散相关蛋白随时间而减少。

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