生物反应器污染监测使用排气的挥发性有机化合物 (VOCs)
Angela L Linderholm1,2, Manohar P Bhandari1,3, Eva Borras3
1Division of Pulmonary, Critical Care and Sleep Medicine, University of California, Davis, Sacramento, CA, USA.
Analytical and bioanalytical chemistry
|December 26, 2024
概括
早期检测细胞培养污染至关重要. 这项研究使用挥发性有机化合物 (VOC) 来比传统方法更快地识别中国仓鼠卵巢 (CHO) 细胞中的微生物污染.
科学领域:
- 生物技术是生物技术.
- 分析化学 分析化学
- 细胞生物学 细胞生物学
背景情况:
- 代谢活跃的细胞释放挥发性有机化合物 (VOC).
- 对细胞培养的实时,非侵入性监测对于健康评估至关重要.
- 细胞培养中的微生物污染会对研究结果产生重大影响.
研究的目的:
- 调整一种利用自然存在的VOC来检测细胞培养中的微生物污染的方法.
- 为了区分中国仓鼠卵巢 (CHO) 细胞与与Staphylococcus epidermidis和Aspergillus fumigatus共同培养的细胞.
- 建立细胞培养污染的早期检测系统.
主要方法:
- 从细胞培养瓶的头部空间中提取VOC,使用涂有聚甲基 (PDMS) 的Twister.
- 提取的挥发性有机物被使用热吸附气体染色学质谱法 (TD-GC-MS) 分析.
- 部分最小平方差异分析 (PLS-DA) 和PLS回归模型用于数据分析.
主要成果:
- 在1-2小时内检测到明显的VOC标志,比传统方法更早地区分受污染的作物.
- PLS-DA模型确定了41种具有高度重要性 (VIP评分>1) 的化合物,用于区分细胞类型和污染物.
- PLS回归预测了微生物细胞度,对S. epidermidis的准确度高 (R2 = 0.891),对A. fumigatus的准确度中等 (R2 = 0.375).
结论:
- 挥发性有机化合物分析提供了一种快速,非侵入性的方法,用于在CHO细胞培养物中早期检测微生物污染.
- 特定的VOC档案可以区分细菌和真菌污染.
- 需要进一步的研究来确定未知的化合物,并阐明它们与受污染的培养物中的代谢变化之间的关系.
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