Biotechnology and Bioprocess Engineering Applications of Raman Spectroscopy

改进的过程分析技术（PAT）的发展是测定水环境中生物分子浓度和构象的一个关键因素。RS可以使用对活细胞无干扰的波长和强度来实现，并且可以与显微镜平台耦合以使用分子振动作为对比机制来提供细胞的无标记化学图像。这些空间分辨的化学图像可以提供关于细胞周期、表型、活力和代谢的丰富信息。最近的技术进步，在RS仪器可在我的实验室进一步允许近实时的时间分辨率的变化，在细胞特性的技术称为受激拉曼光谱（SRS）。该提案描述了利用RS和SRS的三个相关方向的未来研究活动：（1）开发一种非破坏性方法，用于在干细胞衍生的基于细胞的治疗剂的制造过程中表征和监测细胞，这提供了新的QC过程分析技术，增强了传统的QC过程分析技术。（破坏性的）取样细胞分析与原位光学方法，也提供正交分析数据;（2）开发仪器和方法，使血库和医院能够评估与储存有关的降解，从而评估密封储存袋中高价值血液制品的输血适用性;（3）开发分析工具，使生物化学工程师能够更有效地筛选在生物工艺开发过程中为高产细胞生产特定蛋白质的细胞培养物，并更好地了解和监测生产过程中降低生产率的细胞变化。这些是我实验室的新项目的例子，包括一个持续的研究计划的核心，旨在开发有价值的新的光谱工具的基础上RS和SRS的应用在生物技术和生物过程工程。

The development of improved process analytical technologies (PAT) is a crucial Rfer distinct advantages for determining the concentrations and conformations of biomolecules in aqueous environments. RS can be implemented using wavelengths and intensities that are non-perturbing to live cells, and can be coupled with a microscopy platform to provide label-free chemical images of cells using molecular vibrations as a contrast mechanism. These spatially resolved chemical images can provide a wealth of information about cell cycles, phenotypes, viability and metabolism. Recent technological advances in RS instrumentation available in my laboratory further allows near real-time temporal resolution of changes in cellular characteristics using a technique called stimulated Raman spectroscopy (SRS). This proposal describes future research activities in three related directions that exploit both RS and SRS: (1) to develop a non-destructive methodology for characterizing and monitoring cells during the manufacturing of cell-based therapeutics derived from stem cells which provides new QC process analytical technologies that augment conventional (destructive) sampled cell analyses with an in situ optical approach that also offers orthogonal analytical data; (2) to develop instrumentation and methods that allow blood banks and hospitals to assess storage-related degradation, and thus the suitability for transfusion, of high-value blood products within sealed storage bags; (3) to develop analytical tools that allow biochemical engineers to more efficiently screen cultures of cells engineered to produce specific proteins for high-producing cells during bioprocess development, and to better understand and monitor cellular changes during production that degrade productivity. These are examples of new projects in my laboratory that comprise the core of a continuing research program directed toward developing valuable new spectroscopic tools based on RS and SRS for applications in biotechnology and bioprocess engineering.