Cell Technology Group, National Institute for Bioprocessing, Research and Training (NIBRT), Fosters Avenue, Mount Merrion, Blackrock, Dublin, A94 X099, Ireland.
College of Pharmacy, University of Babylon, Babylon, Iraq.
Appl Microbiol Biotechnol. 2021 Apr;105(8):3115-3129. doi: 10.1007/s00253-021-11244-8. Epub 2021 Apr 1.
Four independent mAb-producing CHO cell lines were grown in media supplemented with one of seven protein hydrolysates of animal and plant origin. This generated a 7x4 matrix of replicate cultures which was analysed for viable cell density and mAb productivity. In all cultures, a consistent growth rate was shown in batch culture up to 4 to 5 days. Differences between cultures appeared in the decline phase which was followed up to 7 days beyond the start of the cultures. There was a marginal but significant overall increase (x1.1) in the integral viable cell density (IVCD) in the presence of hydrolysate but a more substantial increase in the cell-specific mAb (qMab) productivity (x1.5). There were individual differences between hydrolysates in terms of enhancement of mAb productivity, the highest being a 166% increase of mAb titre (to 117 mg/L) in batch cultures of CHO-EG2 supplemented with UPcotton hydrolysate. The effect of one of the most active hydrolysates (HP7504) on antibody glycosylation was investigated. This showed no change in the predominant seven glycans produced but a significant increase in the galactosylation and sialylation of some but not all the antibodies. Overall, the animal hydrolysate, Primatone and two cotton-derived hydrolysates provided the most substantial benefit for enhanced productivity. The cotton-based hydrolysates can be viewed as valuable supplements for animal-derived component-free (ADCF) media and as a source for the investigation of chemically defined bioactive components. KEY POINTS: • Protein hydrolysates enhanced both IVCD & qMab; the effect on qMab being consistently greater. • Cotton-based hydrolysates showed high bioactivity and potential for use in serum-free media. • Enhanced galactosylation and sialylation was shown for some of the Mabs tested.
四种独立的产生单抗的 CHO 细胞系在添加了七种动物和植物来源的蛋白质水解物的培养基中生长。这产生了一个 7x4 的重复培养物矩阵，用于分析活细胞密度和单抗生产力。在所有培养物中，在批培养中直到 4 到 5 天，都表现出一致的生长速度。培养物之间的差异出现在衰退阶段，该阶段一直持续到培养物开始后的 7 天。在水解物存在的情况下，整体活细胞密度（IVCD）略有但显著增加（x1.1），但细胞特异性单抗（qMab）生产力增加更多（x1.5）。在水解物增强单抗生产力方面存在个体差异，其中最高的是在补充 UPcotton 水解物的 CHO-EG2 批培养物中，单抗产量增加了 166%（达到 117mg/L）。研究了一种最活跃的水解物（HP7504）对抗体糖基化的影响。这表明产生的主要七种聚糖没有变化，但一些（但不是所有）抗体的半乳糖基化和唾液酸化显著增加。总的来说，动物水解物 Primatone 和两种棉衍生水解物为提高生产力提供了最大的益处。棉基水解物可视为无动物源性成分（ADCF）培养基的有价值补充物，也是研究化学定义生物活性成分的来源。要点：
蛋白质水解物增强了 IVCD 和 qMab；对 qMab 的影响始终更大。
棉基水解物表现出高生物活性，有潜力用于无血清培养基。
一些测试的单抗显示出增强的半乳糖基化和唾液酸化。
