Investigation of Near-Infrared Spectroscopy for Periodic Determination of Glucose in Cell Culture Media in Situ

Abstract

Current techniques used to monitor glucose concentration of cell culture media (CCM) require invasive and tedious handling of the sample for sterile media removal and nutrient replacement. In order to optimize cell culture growth in bioreactors, biosensors must be developed that are capable of monitoring the cell culture processes noninvasively and continuously. In this study, on-line, noninvasive determinations of glucose in cell culture media were investigated via near-infrared spectroscopy (NIR) across the 2.0–2.5 μm combination region. A system was developed, using a unique fiberoptic coupling method and a commercial Fourier transform infrared (FT-IR) spectrometer, to characterize glucose single-beam spectra collected from cell culture media. This novel system is the first of its kind and integrates a completely noninvasive optical probe to measure glucose concentrations within cell culture media, in situ. Spectra recorded from a four-day fibroblast culture with this fiber coupled system and an FT-IR spectrometer have been analyzed and compared with standard clinical chemistry techniques. Partial least-squares (PLS) regression has been used to extract the analyte-dependent information and to build a successful multivariate calibration model. A combination of spectra from cell culture media and prepared media mixtures was used to eliminate unwanted correlations in the calibration data. The combined use of this unique fiberoptic system, PLS, and uncorrelated spectra resulted in a true glucose prediction error of 14.8 mg/dL in an independent validation set.

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

“…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. Thus, a sensor system based on NIR spectroscopy seems to be suitable for RWV monitoring.…”

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

“…The analytical utility of NIR spectroscopy is established for measuring millimolar levels of various biological species in simulated matrices Burmeister et al, 1998;Burmeister and Arnold, 1999) and natural matrices from clinical chemistry (Hazen et al, 1998), fermentation (Cavinato et al, 1990;Ge et al, 1994;Hall et al, 1996;Yano et al, 1998), and cell culture Lewis et al, 2000;McShane and Cote , 1998;Riley et al, 1997Riley et al, , 1998aRiley et al, , 1998bZhou et al, 1995) systems. The chemical information originates from overtone and combination transitions associated with fundamental C±H, O±H, and N±H vibrations.…”

“…Indeed, the well-known relationship between glucose consumption and lactate production generates a concentration correlation that is unacceptable for building PLS calibration models. The implications of cell metabolism on method calibration have been examined in detail elsewhere (Lewis et al, 2000;McShane and Cote , 1998;Riley et al, 1997Riley et al, , 1998aRiley et al, , 1998b. Brie¯y, the calibration samples are best collected at various timepoints during cultivation in order to account for the global chemical changes in the samples.…”

“…Metabolism-induced concentration correlations are created within the samples where glucose and lactate are inversely correlated, glutamine and ammonia are inversely correlated, glucose and glutamine are directly correlated, and lactate and ammonia are directly correlated. The existence of such correlations within the training data set prohibits the generation of an analyte speci®c calibration model Lewis et al, 2000). With concentration correlations in the training data set, accurate analyte measurements cannot be performed independently, but depend on the concentrations of these other components.…”

Near Infrared Spectroscopy in Drug Discovery and Development Processes

“…Calibration models were established for glutamine and glucose with PLS and the models could be used for high concentrations. Studies about cell culture media were made Lewis and co-workers in 2000 [186] and Jung and co-workers in 2002 [187]. In the first study, the authors developed models to predict glucose production using PLS.…”