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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Optimization of lytic phage manufacturing in bioreactor using monolithic supports
Franc Smrekar1, Mateja Ciringer, Janez Jančar
1BIA Separations d.o.o., Ljubljana, Slovenia.
Journal of Separation Science
|June 8, 2011
Summary
Manufacturing large quantities of bacteriophages requires precise cultivation termination to ensure purity. A new chromatographic method accurately estimates phage titer and purity, improving yield and reducing contamination.
Area of Science:
- Microbiology
- Biotechnology
- Bioprocess Engineering
Background:
- Bacteriophage production is crucial for therapeutic applications.
- Achieving high phage quantity and purity is challenging.
- Contamination with host cell DNA reduces phage efficacy.
Purpose of the Study:
- To develop an efficient process for large-scale lytic bacteriophage manufacturing.
- To establish reliable methods for monitoring phage cultivation and ensuring purity.
- To introduce a rapid, on-line method for phage titer estimation.
Main Methods:
- Optimization of bacteriophage cultivation and termination point determination.
- Development of an indirect chromatographic method using methacrylate monoliths for on-line monitoring.
- Implementation of purification strategies including monolithic column chromatography and EDTA treatment.
- Validation of the method using plaque assays and DNA contamination assessment.
Main Results:
- Optimal cultivation termination is critical for high phage quantity and purity.
- The novel chromatographic method accurately correlates with plaque assay results for phage titer.
- Phage fractions exceeding 90% purity were achieved.
- The entire manufacturing and quality control process can be completed within one working day.
Conclusions:
- A robust and efficient protocol for large-scale bacteriophage manufacturing has been established.
- The developed chromatographic method offers a rapid and reliable tool for on-line monitoring and quality control.
- This advancement facilitates the production of high-purity bacteriophages for potential therapeutic use.
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