A Robust HPLC-Mediated Method Development for the Qualitative Monitoring of Water-Soluble Vitamins in Cell Culture Media

Expression of recombinant proteins is highly dependent on the micro-environment provided to the cells during the cell culture process. Typically, the cell culture process at industrial scale lasts for 7–15 days, and maintaining a suitable nutrient-rich micro-environment is extremely critical to achieve the desired quality of the protein. Composition of Media and feed, and their application in the process, is crucial as they decide the quality and quantity of the protein. Most of the commercially available media and feeds come with proprietary label from the supplier, so there is always an uncertainty on the concentration and time of addition during the process, and hence the variability in desired quality is likely, and it is always the case that we are under-feeding few and over-feeding few other nutrients. In general, the media and feeds complex mixture of nutrients, which are required for cell growth, is primarily made up of amino acids, vitamins, trace metals, etc. Understanding the composition of these components in media and their consumption during the cell culture process can significantly help in directing the outcome of the process in a desired manner. In this work, the focus was to make a single high-throughput high-performance liquid chromatography method for profiling of nine water-soluble vitamins (vitamins B and C) with baseline separation and to develop a robust sample preparation procedure to minimize the matrix interference of other components present in the cell culture media. The outcome of the study achieved both objectives, where all nine water-soluble vitamins were obtained as baseline separated, and the same method could detect the presence of vitamins in cell culture components where no baseline drift was observed, which confirmed the development of robust sample preparation.