Amino acid analysis and quantitation is an essential effort for trends and pathways in upstream cell culture media processes to support the adjustment of feed conditions in bioprocessing. This data is used to predict cell biochemical pathways for the production of biologic drugs. Herein, this poster will present improvements in our laboratory that demonstrate a well-known application note from Waters Corporation that has been modified to successfully perform the necessary automated pre-column derivatization for quantifying amino acids in bioreactor samples in fed-batch processes and in chemically defined media used in upstream cell-culture. Specifically, modifications were made to this off-the-shelf method to develop custom standards to thus include six additional amino acids that can be quantified while maintaining resolution, separation, and speed of the method. This pre-column derivatization technique utilizes an amine-derivatizing compound, 6-aminoquinolyl-N-hydroxysuccinimydl carbamate, AQC, to convert both primary and secondary amino acids to stable derivatives. These fluorescently labeled amino acid derivatives are detectable and quantitated according to UV absorbance and fluorescence intensities by using reverse-phase ultra-performance liquid chromatography (UPLC) to produce precise, accurate, and reproducible results. This poster will further articulate the pre-column derivatization methodology essential to optimize automation using Beckman Coulter’s automated liquid handler, Biomek 4000, for approximately sixty samples in under two hours, thus reducing human error and time compared to laborious manual preparation of samples. With a 10.2 minute injection run time, samples can be analyzed overnight to provide quick feedback on decisions for bioreactor feeding strategies and amount based off of amino acid consumption rates. The long-term impact will also be discussed, which will include Janssen’s strategy for optimizing this methodology to increase the sample amount by six-fold, the need for various dilution schemes in expanding the range of media platforms, and future improvements that we aim to employ utilizing automation.<br/><br/>

High throughput automation for quantitation of amino acid analysis via UPLC of basal and feed bioreactor samples in upstream cell culture bioprocesses

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.