Timely process information about fermentation biomass, substrates, intermediates, and products although rarely available, is necessary to make control decisions. NIR, FTIR and Raman spectroscopy offer the opportunity to improve the fermentation processes by incorporating rapid, nondestructive, and multi-constituent analysis of fermentation broth media directly into the fermentation monitoring and control strategies. The objective of our research project is to determine which of the technologies is best for monitoring the cell growth and recombinant protein production of a genetically-modified Escherichia coli strain culture in fed-batch mode. The E. coli strain and the recombinant protein selected is commercially available (Bio-Rad pGLO kit), which produces a green fluorescent protein (GFP) when induced with L-arabinose. We will report on the results from a set of experiments that include: monitoring of fed-batch cultures of E. coli producing recombinant GFP using UV spectrophotometry to monitor cell mass, HPLC to determine the glycerol and acetate concentrations and fluorescence spectrophotometry to monitor GFP production as the primary reference methods to eventually calibrate and build the univariate chemometric models with the NIR, FTIR and Raman spectral data. Further experiments will be conducted to validate the best spectroscopy technique and to build robust models needed to implement them as tools for monitoring cell growth and recombinant protein production. This method would result to be very important to biotechnology industries and the implementation of Process Analytical Technologies (PAT), which has been promoted by the FDA as a preferred mode to produce regulated biotechnology products in the future.

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