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.

中文翻译：

---

代谢组学分析和DOE优化策略，用于提高鸡胚成纤维细胞DF-1细胞的IBDV产量。

由IBD病毒（IBDV）引起的传染性法氏囊病（IBD）具有高度传染性，在鸡胚中接种疫苗已成为预防急性感染的有效手段。然而，目前生产的IBDV疫苗面临严重的批次不稳定和外部污染。鸡胚胎成纤维细胞系DF-1被广泛用于禽病毒的增殖和疫苗的生产。因此，优化DF-1细胞的IBDV生产具有重要的应用价值。结合代谢组学分析和实验设计（DOE）统计策略，这项研究成功地优化了DF-1细胞产生IBDV的过程。通过多变量统计分析对IBDV感染和未感染DF-1细胞中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，磷酸戊糖途径，核苷合成途径，脂质代谢和谷胱甘肽代谢的细胞内代谢强度被上调，而TCA循环则被轻微下调。根据代谢组学结果和DOE统计优化方法，确定适合IBDV增殖的添加剂组分。在对照培养基中外源添加半胱氨酸，蛋氨酸，赖氨酸和核苷后，IBDV滴度增加了20.7倍，这与多元二次方程的预测结果（20.0倍）相符。这项研究提供了有效生产IBDV疫苗的策略，并有可能被用于改善其他病毒疫苗和生物制剂的生产。