Nondestructive near‐infrared spectroscopic measurement of multiple analytes in undiluted samples of serum‐based cell culture media

Abstract

An adaptive calibration procedure is used to build selective multivariate calibration models for the measurement of glucose, lactate, glutamine, and ammonia in undiluted serum-based cell culture media. This adaptive procedure removes metabolism-induced covariance between these analytes in a series of calibration samples collected during the cultivation of PC-3 human prostate cancer cells. Partial least-squares calibration models are generated from single-beam near-infrared (NIR) spectra collected over the 4800-to 4200-cm )1 combination spectral range. Calibration models were generated with both the full spectral range and optimized spectral ranges. In both cases, the number of model factors was optimized and model validity was determined by comparing analyte concentrations predicted from a series of independent and unaltered samples that were obtained during a subsequent cultivation of the PC-3 cells. Similar analytical performance was achieved with fewer model factors when the optimized spectral range was used. The lowest standard errors of prediction were 0.82, 0.94, 0.55, and 0.76 mM for glucose, lactate, glutamine, and ammonia, respectively. Different spectral ranges were optimal for each analyte and the optimized spectral range coincided with the distinguishing spectral features of the analyte. The results of this study demonstrate that NIR spectroscopy can be used effectively in the off-line measurement of important nutrients (glucose and glutamine) and byproducts (lactate and ammonia) in a serum-based animal cell culture medium.

Automated Classification of Bioprocess Based on Optimum Compromise Whitening and Clustering

“…The model was successfully able to detect abnormal process conditions, which resulted from three known fault types, namely irregular thermal heating, elevated dissolved oxygen values, and large variation in agitation 3 . PLS calibration models of NIR spectra have been utilized for the measurement of glucose, lactate, glutamine, and ammonia in undiluted serum-based cell culture media 4 . Robust, analyte-specific models were generated, and the low values of standard errors of prediction for each analyte demonstrate that the models can be used to (off-line) determine the important nutrient and byproduct content in a serum-based cell culture medium.…”

On‐line monitoring of human prostate cancer cells in a perfusion rotating wall vessel by near‐infrared spectroscopy

“…Simultaneous measurement eliminates the need for multiple sensors, and noninvasive measurement avoids the need for elaborate sterilizations, such as, for example, with the amperometric sensor described by Xu et al (2002). Rhiel et al (2002a) generated calibration models able to monitor glucose, lactate, glutamine, and ammonia with SEPs of 0.82, 0.94, 0.55, and 0.76 mM, respectively (Rhiel et al, 2002a). Similar results were obtained by Zhou et al (1995), Chung et al (1996), Riley et al (1997Riley et al ( , 1998aRiley et al ( , 1998b, McShane and Coté (1998), and Lewis et al (2000) for cell cultures, and Cavinato et al (1990), Ge et al (1994), Hall et al (1996), and Yano et al (1998) for fermentations.…”

“…For this reason, Rhiel et al (2002a) investigated whether four key metabolites (glucose, lactate, glutamine, and ammonia) could be monitored with near-infrared (NIR) spectroscopy. The advantage of spectroscopic sensors is that multiple components can be measured simultaneously and noninvasively (Rhiel et al, 2002a). Simultaneous measurement eliminates the need for multiple sensors, and noninvasive measurement avoids the need for elaborate sterilizations, such as, for example, with the amperometric sensor described by Xu et al (2002).…”

“…In addition, the waste products of the cellular metabolism are also important as they may accumulate to toxic levels when the nutrient concentration is not controlled (Glacken et al, 1986). For this reason, Rhiel et al (2002a) investigated whether four key metabolites (glucose, lactate, glutamine, and ammonia) could be monitored with near-infrared (NIR) spectroscopy. The advantage of spectroscopic sensors is that multiple components can be measured simultaneously and noninvasively (Rhiel et al, 2002a).…”

“…This adaptive calibration approach yields more robust calibration models (Rhiel et al, 2002c) and was used as a basis in this work. In fact, basic calibration model development has been described previously (Rhiel et al, 2002a).…”