Cell culture media: NMR approaches to evaluating quality and nutrient consumption

Cell culture media is widely used to provide nutrients for cell growth in research, diagnostic and manufacturing applications [1]. Use in research and manufacturing applications to produce secondary metabolites from microbes is long standing [2]. Cell culture media use is increasing as a result of the growing focus on the development and production of biologics and biosimilars in the pharmaceutical industry. Typical cell culture media contain a mixture of defined nutrients that are expected to produce the desired product. The nutrients are often small molecule components that may be monitored using NMR spectroscopy. In this work we present techniques that were utilized to enhance the analysis of cell culture media. The ability for automated NMR analysis for quality control was evaluated. Solvent suppression and region selective 1D and 2D homonuclear and heteronuclear NMR techniques enhanced the monitoring of nutrients consumed and characterization of metabolites produced in the media. Drift compensation features of modern NMR systems provided the ability to obtain spectra of the pure media without requiring addition of a deuterium lock solvent, allowing for real time NMR monitoring of the media. These studies evaluate the ability to monitor fermentations in either (1) batch mode, sampling aliquots at time points and submitting them for analysis, or (2) continuous monitoring in flow mode (reaction monitor). NMR monitoring may allow optimization of real time conditions of the fermentation for maximum production of a desired product.

Keywords: NMR, cell culture media, reaction monitoring.

References:

[1] Vijaya SN, Udayasri PV, Aswani KY, Ravi BB, Phani KY, Vijay VM. Advancements in the production of secondary metabolites. J Nat Prod 2010; 3: 112 – 123

[2] Hussain MS, Fareed S, Ansari S, Rahman MA, Ahmad IZ, Saeed M. Current approaches toward production of secondary plant metabolites. J Pharm Bioallied Sci 2012; 4: 10 – 20