Historically, drugs, medications and other pharmaceuticals have been the domain of synthetic organic chemists. These specialized chemicals have typically been designed in the laboratory then transferred to large scale production by chemical engineers. Yet, a growing number of medically important molecules, including proteins, hormones, and some antibiotics, are simply just too large and complex to easily synthesize in the lab, much less on a large industrial scale. Pharmaceutical companies are increasingly turning towards biological systems for large scale production of these elaborate substances. Cultures for these syntheses often use genetically modified bacteria, yeast, insect, or mammal cells. The cultures are placed in large tanks under carefully controlled conditions that are conducive to cell growth and target molecule production. To facilitate optimal production of the molecule of interest, the culture media must be monitored and adjusted with respect to nutrients, waste products, temperature, pH, and other parameters. Fourier Transform Near-Infrared (FT-NIR) spectroscopy is an excellent method of monitoring these crucial parameters. FT-NIR is capable of spectroscopically analyzing cell cultures in situ, yielding real-time results for multiple components in the media. These components include nutrients such as glucose; waste materials such as lactic acid and ammonia; and target protein. Additionally, other important culture parameters such as pH and cell density can be monitored. Because the analysis is spectroscopic in nature, samples need not be withdrawn for offline analysis, which saves considerable time and circumvents contamination issues. Furthermore, the Antaris line of FT-NIR analyzers includes internal calibration standards which allow instrument qualification without having to stop the process or remove the probe from the culture media. Below is a summary of a typical pilot project for analyzing and tracking a cell culture used to manufacture a recombinant protein. | Design of ExperimentInstrument: Antaris II Method Development Sampling (MDS) FT-NIR analyzer with heated transmission cell module
Technique: Transmission analysis in a temperature-controlled cell.
Rationale: Real-time analysis would provide superior results and tracking of the cell culture health and production of the material of interest. Other methods, such as off-line sampling require stopping the process and are time consuming. Nutrients, waste products and other parameters can be analyzed spectroscopically with the Antaris II FT-NIR analyzer.
Samples: Human embryonic kidney cells (HEK 293) were used in chemically-defined media and allowed to grow for 10 days. The media was spectroscopically analyzed with the Antaris system throughout the process. Partial Least Squares (PLS) analysis was performed on the first-derivative spectra. The nutrients glucose and glutamine, as well as the waste product ammonia were accurately measured and tracked well with expected results. Additionally pH and viable cell density were accurately predicted, indicating FT-NIR is a highly valuable tool for monitoring cell cultures in biopharmaceutical manufacturing. |
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