UV resonance Raman spectroscopy: a process analytical tool for host cell DNA and RNA dynamics in mammalian cell lines
UV resonance Raman spectroscopy: a process analytical tool for host cell DNA and RNA dynamics in mammalian cell lines
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紫外共振拉曼光谱：哺乳动物细胞系宿主细胞 DNA 和 RNA 动力学的过程分析工具
DOI：
10.1002/jctb.4420
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发表时间：
2014
影响因子：
3.4
通讯作者：
Ashton L
中科院分区：
文献类型：
--
作者：
Ashton L
背景拉曼光谱学的最新进展导致了快速原位拉曼探针的开发，其可以识别并允许评估哺乳动物细胞培养物中复杂成分的质量。一种特定的拉曼技术，即紫外共振拉曼 (UVRR) 光谱，有潜力作为哺乳动物细胞培养基中残留细胞 DNA 和 RNA 的探针。 结果 鉴定出中等细胞足迹样品的 DNA 和 RNA UVRR 光谱曲线的变化，这些变化与收获时间和细胞裂解增加有关，而细胞裂解则与细胞活力丧失相关。在丁酸钠处理的细胞培养上清液中还观察到 DNA 和 RNA 增加。下游生物加工过程中初级和二级澄清方法导致的 DNA 和 RNA 谱的变化也可以通过 UVRR 光谱来确定。结论本研究证明了 UVRR 光谱作为监测可能污染细胞培养基的残留 DNA 和 RNA 变化的工具的实用性。该技术的应用有可能改进回收技术并保证 DNA/RNA 可靠清除到可接受的安全水平。 © 2014 中国化工学会
BACKGROUNDRecent advances in Raman spectroscopy have resulted in the development of rapid,in situRaman probes that can identify and allow the assessment of the quality of complex constituents in mammalian cell culture. One specific Raman technique, UV resonance Raman (UVRR) spectroscopy, has potential as a probe for residual cellular DNA and RNA in mammalian cell culture medium.RESULTSVariations in DNA and RNA UVRR spectral profiles of medium‐cellular footprint samples were identified and related to time of harvest and increased cell lysis that is associated with a loss in cell viability. Increased DNA and RNA were also observed in the cell culture supernatant in response to sodium butyrate treatment. Variation in DNA and RNA profiles as a result of both primary and secondary clarification methods during downstream bioprocessing could also be determined with UVRR spectroscopy.CONCLUSIONSThis study has demonstrated the utility of UVRR spectroscopy as a tool to monitor variations in residual DNA and RNA that may contaminate cell culture medium. Application of this technique has the potential for both improvement of recovery techniques and assurance of reliable clearance of DNA/RNA to acceptable safety levels. © 2014 Society of Chemical Industry