The use of flow cytometry to detect nucleic acids attached to the surface of Escherichia coli in high cell density fed-batch processes

• E. coli was grown in an aerobic fed-batch process for the production of a recombinant protein (rhGH). The cells were examined by flow cytometry and PI (propidium iodide) staining. The fluorescence of the PI-stained cells increased with increasing concentrations of DNA in the medium. Furthermore, DNA and RNA attached to the cell could partly be degraded with DNase/RNase and the fluorescence decreased. Formate excretion during the aerobic processes may be due to DNA and possibly also RNA attached to the cell surface, so creating diffusion resistance.

Nyckelord

DNA release

E. coli

flow cytometry

formate formation

culture

growth

w3110

flux

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art (ämneskategori)