Optimisation of Cell Growth and Recombinant Protein Production in Small-Scale Culture Vessels by Using Optical Sensors for On-line Measurement of Dissolved Oxygen

Jäger, Volker; Apati, Giannini Paszinick; Konisch, Nadine

doi:10.1007/978-94-007-0884-6_61

Cited by 1 publication

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“…Another approach is the optical measurement of dissolved oxygen tension inside a culture broth. Thereby, an oxygen sensitive fluorophore spot is attached at the inside of a shake flask . Since fluorophore spots can be very small in size, this method is transferable to microtiter plates .…”

Section: Introductionmentioning

confidence: 99%

“…[5][6][7] Although monitoring of oxygen supply in fermenters using oxygen probes is a long-standing common practice 8,9 ; it is rather new in small-scale bioreactors like shake flasks and microtiter plates. [10][11][12][13][14][15][16] Anderlei et al presented the so-called Respiration Activity MOnitoring System (RAMOS)technology to monitor the OTR, carbon dioxide transfer rate and the respiratory quotient within eight parallel shake flasks. 10 This is a noninvasive method based on electrochemical partial pressure measurements in the headspace of shake flasks.…”

Section: Introductionmentioning

confidence: 99%

“…Thereby, an oxygen sensitive fluorophore spot is attached at the inside of a shake flask. 11 Since fluorophore spots can be very small in size, this method is transferable to microtiter plates. 12 An alternative method for both, shake flask and microtiter plate, represents the application of infrared fluorescent nanoparticles within the culture broth.…”

Section: Introductionmentioning

confidence: 99%

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Prediction of recombinant protein production by Escherichia coli derived online from indicators of metabolic burden

Mühlmann

Forsten

Noack

et al. 2018

Biotechnology Progress

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Background: The oxygen transfer rate (OTR) and the biomass concentration are two important parameters describing a microbial fermentation. It has been shown before that from the course of these parameters over time information on metabolic burden during heterologous protein production can be obtained. While online monitoring in large fermenters is ubiquitously established, it is still not a common practice in small‐scale cultures. Nevertheless, several techniques like the Respiration Activity MOnitoring System (RAMOS) device for online monitoring of the OTR in shake flasks and the BioLector device for measuring scattered light (ScL) representing biomass in microtiter plates have been developed. Results: A new microtiter plate‐based method is presented that reveals how online derived ScL signals can be transformed into signals that are proportional to the courses of OTR over time for Escherichia coli. The transformed signal is obtained by simply taking the first derivative of ScL (dScL/dt). The proportionality of both parameters is successfully validated for the strains E. coli BL21(DE3) and Tuner(DE3) expressing cellulases and the fluorescent protein FbFP, respectively. Relative amounts of produced heterologous proteins are predicted exclusively based on the course of the transformed ScL signal. A variety of induction conditions with varying inducer concentration and induction time were investigated with this method. Conclusion: The presented method based on ScL measurement allows for high‐throughput online determination of signals proportional to OTR courses. They enable the interpretation of physiological states and offer the possibility to predict the recombinant protein production in E. coli. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1543–1552, 2018

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Section: Introductionmentioning

confidence: 99%