Short Lecture : Monitoring nutrients during process development
Near-infrared spectroscopy is considered to be one of the most promising spectroscopic techniques for upstream bioprocess monitoring and control. Traditionally the nature of near-infrared spectroscopy has demanded multivariate calibration models to relate spectral variance to analyte concentrations. The resulting analytical measurements have proven unreliable for the measurement of metabolic substrates for bioprocess batches performed outside the calibration process. This paper presents results of an innovative near-infrared spectroscopic monitor designed to follow the concentrations of glycerol and methanol, as well as biomass, in real time and continuously during the production of a monoclonal antibody by a Pichia pastoris high cell density process. A solid-state instrumental design overcomes the ruggedness limitations of conventional interferometer-based spectrometers. Accurate monitoring of glycerol, methanol, and biomass is demonstrated over 274 days post-calibration. In addition, the first example of feedback control to maintain constant methanol concentrations, as low as 1 g/L, is presented. Post-calibration measurements over a nine-month period illustrate a level of reliability and robustness that promises its adoption for on-line bioprocess monitoring throughout product development, from early laboratory research and development to pilot and manufacturing scale operation.