Biotechnology and Bioprocess Engineering Applications of Raman Spectroscopy

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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.**