在本研究中,使用12种细胞培养基进行初步筛选,选择性能最好的四种培养基进行进一步研究。使用增强单纯形质心混合物设计评估了用于YFV生产的四种介质混合物的优化。在所有被调查的不同模型中,二次模型被认为是探索混合设计的最合适模型。发现M10对YFV生产的影响最大,其次是M9、M4和M1。与它们与其他介质的共混物相比,M1和M4介质的单独利用产生更高的收益。当M1培养基与其他培养基组合时,YFV滴度降低。与它们各自的浓度相比,M9和M10培养基的组合使用导致更高的病毒产量。发现从初级CEF获得较高滴度的YFV的最佳比率为约38:62,其中M9和M10是最有利的培养基混合物。培养基混合物的使用导致病毒滴度的显着增加高达2.6×108PFU/ml或2log滴度产量,相当于1.92×105剂量,生长条件或其他过程因素没有任何变化。这项研究得出的结论是,可以有效地利用混合物设计来选择培养基混合物的最佳组合,以提高细胞培养物中的病毒产量。
In the present study, an initial screening was conducted using 12 types of cell culture media, and four media with the best performance were selected for further study. The optimization of four media blend for YFV production was evaluated using an Augmented simplex centroid mixture design. Among all the different models that were investigated, the quadratic model was found to be the most appropriate model for exploring mixture design. It was found that M10 exhibited the greatest impact on YFV production, followed by M9, M4, and M1. The utilization of M1 and M4 media individually yielded higher compared to their blends with other media. The YFV titers were reduced when M1 media was combined with other media. The utilization of M9 and M10 media in combination resulted a higher viral yield compared to their respective concentrations. The optimal ratio for achieving a higher titer of YFV from primary CEFs was found to be approximately 38:62, with M9 and M10 being the most favorable media blend. The use of a media mixture led to a significant increase of virus titer up to 2.6 × 108 PFU/ml or 2 log titer yield, which is equivalent to 1.92 × 105 doses, without any changes to growth conditions or other process factors. This study concluded that the utilization of a mixture design could be efficiently employed to choose the optimal combination of media blends for enhanced viral production from cell culture.