Application of Process Raman Spectroscopy for bioprocessing using gene edited production hosts and cell-free systems

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Raman spectroscopy (RS) is a non-destructive vibrational spectroscopy technique that provides sharp spectral features that correlate to a sample's chemical or molecular structure in aqueous solution. Raman measurements of biological molecules, cells and tissues have been well established with many biomedical applications focused on disease detection and in vivo glucose monitoring. In parallel there has been substantial developments in the use of RS to monitor fermentation processes. In situ, simultaneous measurement of nutrients, metabolites or by-products, cell density (or biomass), have made RS an important Process Analytical Technology in industrial bioprocessing. A number of global biomanufacturers including for example Lonza, GSK, Merck Novartis Hoffman La Roche have already implemented online Raman Spectroscopy for process control and optimisation of biomanufacturing. Whilst genome editing has opened up opportunities in gene and cell therapy applications, it can also be used in bioprocessing to improve cell growth characteristics; cell productivity and product titre and optimise and/or improve product quality; improve cell free extracts as production hosts. An alternative to cell culture for bioprocessing is the use of cell-free protein expression (CPE) systems based on cell lysates. Recently CPE has been demonstrated as a potential production host for a variety of biologics, vaccines and antimicrobials. The advantages of CPE systems for bioprocessing it that they are non-living and not prone to contamination, have semi-defined components and an assay format that offer flexibility in adjusting the reaction mix and expression process. The overall aim of the proposed project is to explore the application of RS into new areas such as genome edited cell cultures and cell-free systems with an industrial collaborator Kaiser (Endres-Hauser group) who design and manufacture Raman probes. The proposed project brings together the expertise of Freemont group in CPE and cell culture, the Kazarian group in RS and London Biofoundry in synthetic biology tool development.