Biotechnology and Bioprocess Engineering Applications of Raman Spectroscopy

改进的过程分析技术（PAT）的发展对于确定水环境中生物分子的浓度和构象具有重要的优势。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结合使用分子振动作为对比机制来提供细胞的无标签化学图像。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。在我的实验室中，RS仪器的最新技术进步进一步允许使用一种称为受激拉曼光谱（SRS）的技术对细胞特性变化的近实时时间分辨率进行分析。本提案描述了利用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，用于在干细胞为基础的治疗药物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，通过原位光学方法增强传统的（破坏性）取样细胞分析，并提供正交分析数据；(2)开发仪器和方法，使血库和医院能够评估密封储存袋中高价值血液制品的储存相关降解，从而评估其是否适合输血；(3)开发分析工具，使生化工程师能够更有效地筛选培养的细胞，以便在生物过程开发过程中为高产细胞生产特定蛋白质，并更好地了解和监测生产过程中降低生产力的细胞变化。这些是我的实验室新项目的例子，它们构成了一个持续研究计划的核心，该计划旨在开发基于RS和SRS的有价值的新光谱工具，用于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.