A challenging aspect with the use of protein hydrolysates in commercial manufacturing processes of recombinant therapeutic proteins is their impacts on the protein production due to a lack of understanding of batch-to-batch variability. Soy hydrolysates variability and its impact on fed-batch production of a recombinant monoclonal antibody (mAb) expressed in Sp2/0 cells were studied using 37 batches from the same vendor. The batch-to-batch variability of soy hydrolysates impacted cell growth, titer and product quality. Physicochemical characterization of batches confirmed that soy hydrolysates are mainly a source of amino acids and peptides containing lower amounts of other components such as carbohydrates and chemical elements in cell culture media. Soy hydrolysates composition of different batches was consistent except for trace elements. Statistical analyses identified iron as a potential marker of a poor process performance. To verify this correlation, two forms of iron, ferric ammonium citrate and ferrous sulfate, were added to a batch of soy hydrolysates associated to a low level of iron during cell culture. Both forms of iron reduced significantly cell growth, mAb titer and increased level of the acidic charge variants of the mAb. Consequently, trace element composition of soy hydrolysates or of all incoming raw materials might lead to significant impacts on process performance and product quality and therefore need to be tightly controlled.

中文翻译：

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大豆蛋白水解物的表征及其铁含量对鼠杂交瘤细胞系产生单克隆抗体的影响

在重组治疗性蛋白质的商业生产过程中使用蛋白质水解物的一个具有挑战性的方面是，由于缺乏对批次间变异性的了解，它们对蛋白质生产的影响。使用来自同一供应商的 37 个批次研究了大豆水解物的变异性及其对在 Sp2/0 细胞中表达的重组单克隆抗体 (mAb) 的补料分批生产的影响。大豆水解物的批次间差异会影响细胞生长、滴度和产品质量。批次的物理化学特性证实，大豆水解物主要是氨基酸和肽的来源，在细胞培养基中含有较少量的其他成分，如碳水化合物和化学元素。除微量元素外，不同批次的大豆水解物组成一致。统计分析确定铁是工艺性能不佳的潜在标志。为了验证这种相关性，将两种形式的铁（柠檬酸铁铵和硫酸亚铁）添加到一批在细胞培养过程中与低水平铁相关的大豆水解物中。两种形式的铁都显着降低了细胞生长、mAb 滴度并增加了 mAb 酸性电荷变体的水平。因此，大豆水解产物或所有进入原材料的微量元素组成可能会对工艺性能和产品质量产生重大影响，因此需要严格控制。在细胞培养过程中，将其添加到与低水平铁相关的一批大豆水解物中。两种形式的铁都显着降低了细胞生长、mAb 滴度并增加了 mAb 酸性电荷变体的水平。因此，大豆水解产物或所有进入原材料的微量元素组成可能会对工艺性能和产品质量产生重大影响，因此需要严格控制。在细胞培养过程中，将其添加到与低水平铁相关的一批大豆水解物中。两种形式的铁都显着降低了细胞生长、mAb 滴度并增加了 mAb 酸性电荷变体的水平。因此，大豆水解产物或所有进入原材料的微量元素组成可能会对工艺性能和产品质量产生重大影响，因此需要严格控制。