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Intracellular monitoring of superoxide dismutase expression in an Escherichia coli fed-batch cultivation using on-line disruption with at-line surface plasmon resonance detection by Igor Vostiar; Jan Tkac; Carl-Fredrik Mandenius is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Intracellular monitoring of superoxide dismutase expression in an Escherichia coli fed-batch cultivation using on-line disruption with at-line surface plasmon resonance detection about?

An on-line cell disruption system for at-line monitoring of the intracellular concentration of recombinant human superoxide dismutase (rhSOD) in a genetically modiWed Escherichia coli strain, HMS174(DE3) (pET11a/rhSOD), in bioreactor cultivations is described. The sampled bacteria were disrupted on-line by rapid mixing with a nonionic detergent. The recombinant protein content of the lysed bacterial sample was quantitated by a subsequent surface plasmon resonance biosensor with a speciWc monoclonal antibody. Extraction eYciency of the monitoring system was optimized with respect to the Xow rate ratio of the cell suspension and the detergent at relevant cell densities with the aim to attain rapid monitoring. Monitoring was demonstrated for a shake Xask culture and a glucose-limited fed-batch cultivation. The results are compared with a traditional enzyme-linked immunosorbent assay method showing a correlation coeYcient of R 2 D 0.97. Extraction eYciency of rhSOD reached 95-99% at a total processing time of 1.8-2.6 min and a contact time of 0.8-1.4 min. The possibility of extending the monitoring system to other intracellular proteins is discussed.

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Author

Igor Vostiar; Jan Tkac; Carl-Fredrik Mandenius

Publisher

Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)

Published

2005

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)