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Simultaneous measurement of glucose and glutamine in insect cell culture media by near infrared spectroscopy

✍ Scribed by Mark R. Riley; Martin Rhiel; Xiangji Zhou; Mark A. Arnold; David W. Murhammer


Publisher
John Wiley and Sons
Year
1997
Tongue
English
Weight
136 KB
Volume
55
Category
Article
ISSN
0006-3592

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✦ Synopsis


The purpose of this study was to develop noninvasive techniques to monitor the composition of cell culture media in insect cell bioreactors. Such a monitor could be used in conjunction with a fed-batch feeding scheme to ensure that cells are maintained in an optimal environment for growth and protein production. Glucose and glutamine concentrations in an insect cell culture bioreactor were determined off-line with near-infrared (NIR) absorption spectroscopy. Spectra were collected from 5000 to 4000 cm -1 with a 1.5-mm optical path length. Partial least squares (PLS) regression was applied to correlate the collected spectra with the concentration of the desired analytes. Under the culture conditions evaluated here, glucose and glutamine concentrations ranged from 38 to 55 mM and from 3 to 13 mM, respectively. Accurate measurements of glucose and glutamine in insect cell culture samples were possible over these entire ranges. The standard error of prediction (SEP) and mean percent error (MPE) for glutamine were 0.52 mM and 5.3%, respectively. Glucose could be measured with an SEP of 1.30 mM and an MPE of 2.3%. These levels of error are quite low considering the changing complexity of the growth media due to the shifting levels of amino acids, carbohydrates, yeastolate, proteins, and cell debris. This study represents an important step in the development of noninvasive on-line monitoring devices for cell culture bioreactors.


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Nondestructive near-infrared spectroscop
✍ Martin Rhiel; Michael B. Cohen; David W. Murhammer; Mark A. Arnold 📂 Article 📅 2001 🏛 John Wiley and Sons 🌐 English ⚖ 170 KB

## 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