State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology (ECUST), Shanghai, People's Republic of China.
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology (ECUST), Shanghai, People's Republic of China.
J Biotechnol. 2020 Jan 10;307:114-124. doi: 10.1016/j.jbiotec.2019.10.018. Epub 2019 Nov 4.
Infectious bursal disease (IBD) caused by IBD virus (IBDV) is highly contagious viral and vaccination in chicken embryo has been an effective mean to prevent acute infection. However, the current production of IBDV vaccine faces serious batch instability and external contamination. The chicken embryonic fibroblast cell line DF-1 is widely used for the proliferation of avian viruses and vaccine production. Thus, optimizing the production of IBDV by DF-1 cells has an important application value. Combining metabolomics analysis and a Design of Experiments (DOE) statistical strategy, this study successfully optimized the process of IBDV production by DF-1 cells. Differential analysis and time series analysis of metabolite data in both IBDV-infected and uninfected DF-1 cells were performed by multivariate statistical analysis. The results showed that the intracellular metabolite intensities of glycolysis, the pentose phosphate pathway, the nucleoside synthesis pathway, lipid metabolism, and glutathione metabolism were upregulated, and the TCA cycle underwent a slight downregulation after IBDV infection of DF-1 cells. Based on the metabolome results and DOE statistical optimization method, the additive components suitable for IBDV proliferation were determined. The IBDV titer increased by 20.7 times upon exogenous addition of cysteine, methionine, lysine and nucleosides in the control medium, which is consistent with the predicted result (20.0 times) by a multivariate quadratic equation. This study provides a strategy for the efficient production of IBDV vaccines and could potentially be utilized to improve the production of other viral vaccines and biologics.
传染性囊病（IBD）由传染性囊病病毒（IBDV）引起，具有高度传染性，鸡胚接种疫苗是预防急性感染的有效手段。然而，目前 IBDV 疫苗的生产面临着严重的批次不稳定和外部污染问题。鸡胚成纤维细胞系 DF-1 广泛用于禽类病毒的增殖和疫苗生产。因此，优化 DF-1 细胞中 IBDV 的生产具有重要的应用价值。本研究结合代谢组学分析和实验设计（DOE）统计策略，成功优化了 DF-1 细胞中 IBDV 的生产过程。通过多元统计分析对 IBDV 感染和未感染的 DF-1 细胞中的代谢物数据进行差异分析和时间序列分析。结果表明，IBDV 感染 DF-1 细胞后，糖酵解、戊糖磷酸途径、核苷合成途径、脂代谢和谷胱甘肽代谢的细胞内代谢物强度上调，三羧酸循环略有下调。基于代谢组学结果和 DOE 统计优化方法，确定了适合 IBDV 增殖的添加剂成分。在对照培养基中外源添加半胱氨酸、蛋氨酸、赖氨酸和核苷可使 IBDV 滴度提高 20.7 倍，与多元二次方程的预测结果（20.0 倍）一致。本研究为 IBDV 疫苗的高效生产提供了一种策略，有望用于提高其他病毒疫苗和生物制剂的生产效率。
