Journal article
On-line near infrared bioreactor monitoring of cell density and concentrations of glucose and lactate during insect cell cultivation
Journal of biotechnology, Vol.173(1), pp.106-111
03/10/2014
DOI: 10.1016/j.jbiotec.2014.01.009
PMID: 24452098
Abstract
•Online near-infrared bioreactor monitoring was demonstrated during insect cell cultivation.•Raw culture medium was circulated through an autoclavable NIR cell to a bioreactor in a closed-loop system.•Both cell density and concentrations of substrate and by-product were extracted from one NIR spectrum. Near infrared spectroscopy is demonstrated as a suitable method for monitoring real time cell density and concentrations of glucose and lactate during insect cell cultivation. The utility of this approach is illustrated during the cultivation of Trichoplusia ni BTI-Tn-5B1-4 insect cells in a stirred-tank bioreactor. On-line near infrared measurements are made by passing unaltered culture medium through an autoclavable near infrared flow-through sample cell during the cultivation process. Single-beam near infrared spectra were collected over the combination spectral range (5000–4000cm−1) through a 1.5mm path length sample. Cell density calibration model was established by uni-variable linear regressions with measured mean absorbance values of on-line spectra collected during a cultivation run. Calibration models are generated for glucose and lactate by regression analysis of both off line and on line spectra collected during a series of pre-measurement cultivation runs. Analyte-specific calibration models are generated by using a combination of spectra from both natural, unaltered samples and samples spiked with known levels of glucose and lactate. Spiked samples are used to destroy concentration correlations between solutes, thereby enhancing the selectivity of the calibration models. Absorbance spectra are used to build partial least squares calibration models for glucose and lactate. The calibration model for cell density corresponds to a univariate linear regression calibration model based on the mean absorbance between 4750 and 4250cm−1. The standard errors of prediction are 1.54mM, 0.83mM, and 0.38×106cells/mL for the glucose, lactate, and cell density models, respectively.
Details
- Title: Subtitle
- On-line near infrared bioreactor monitoring of cell density and concentrations of glucose and lactate during insect cell cultivation
- Creators
- Jiang Qiu - Department of Chemical and Biochemical Engineering, Optical Science and Technology Center, University of Iowa, Iowa City, IA 52242, United StatesMark A Arnold - Department of Chemistry, Optical Science and Technology Center, University of Iowa, Iowa City, IA 52242, United StatesDavid W Murhammer - Department of Chemical and Biochemical Engineering, Optical Science and Technology Center, University of Iowa, Iowa City, IA 52242, United States
- Resource Type
- Journal article
- Publication Details
- Journal of biotechnology, Vol.173(1), pp.106-111
- Publisher
- Elsevier B.V
- DOI
- 10.1016/j.jbiotec.2014.01.009
- PMID
- 24452098
- ISSN
- 0168-1656
- eISSN
- 1873-4863
- Grant note
- name: Microgravity Science & Applications Division of the National Aeronautics and Space Administration, award: NAG8-1592
- Language
- English
- Date published
- 03/10/2014
- Academic Unit
- Center for Biocatalysis and Bioprocessing; Fraternal Order of Eagles Diabetes Research Center; Chemistry; Chemical and Biochemical Engineering
- Record Identifier
- 9983985842602771
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