A Lack of Inhibitory Action of Bacteriophage T4 Ghosts in the presence of EDTA

Okamoto, Koji; Yonemoto, Rieko

doi:10.1099/0022-1317-30-1-141

Cited by 1 publication

(1 citation statement)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While EDTA seems to be a simple and inexpensive solution to prevent DNA degradation making in this way process more robust, one has to take into account that presence of the EDTA can also inhibit phage growth 27. Because of that it should not be added at the beginning of fermentation but only when the final phage titer is achieved.…”

Section: Resultsmentioning

confidence: 99%

Optimization of lytic phage manufacturing in bioreactor using monolithic supports

Smrekar

Ciringer

Jančar

et al. 2011

J of Separation Science

View full text Add to dashboard Cite

A process for manufacturing large quantities of lytic bacteriophages was developed. Determination of cultivation termination was found to be essential to achieve high phage quantity and purity. When optimal cultivation termination is missed, phage fraction was found to be highly contaminated with deoxyribonucleic acid released from Escherichia coli cells. Besides, an already established method for monitoring of phage cultivation based on optical density, where its peak indicates point when maximal phage titer is achieved, a new indirect chromatographic method using methacrylate monoliths is proposed for on-line estimation of phage titer. It is based on the measurement of released E. coli deoxyribonucleic acid and shows high correlation with phage titer obtained from plaque assay. Its main advantage is that the information is obtained within few minutes. In addition, the same method can also be used to determine purity of a final phage fraction. Two strategies to obtain highly pure phage fractions are proposed: an immediate purification of phage lysate using monolithic columns or an addition of EDTA before chromatographic purification. The developed protocol was shown to give phage purity above 90% and it is completed within one working day including cultivation and phage titer in the final formulation using developed chromatographic method.

show abstract

Section: Resultsmentioning

confidence: 99%