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One-step salting-out extraction of bacteriophage from its infection broth of Acinetobacter baumannii
Suyang Duan1, Yuesheng Dong1, Yongping Xu1
1School of Bioengineering, Dalian University of Technology, Dalian 116024, Liaoning, China.
Abstract:
Phages as a potential alternative antibiotic for multidrug-resistant bacterial infections are receiving great attention worldwide. However, the traditional separation and purification of phage are cumbersome, time-consuming, costly and inefficient. In this study, phage phiAB9 for multidrug-resistant Acinetobacter baumannii was separated and purified by a simple and cost-saving one-step salting-out extraction (SOE). Several kinds of salts and organic solvents without effect on phage survival were chosen to form the SOE systems, and the composition of SOE systems were optimized according to the phage recovery rate and impurity removal rate. After one-step SOE by an optimal system composed of 18% (w/w) ammonium citrate and 40% (w/w) ethyl acetate, the recovery rate of phage in the middle phase could reach to 90.82%, and most proteins (99.57%), cells (97.98%) and endotoxin (84.08%) were well removed, with a concentration factor of 210 and the purification factors of phage to proteins, cells and endotoxin were 303.64, 133.55 and 5.36, respectively. Comparing with two-step SOE and traditional aqueous two-phase extraction, one-step SOE may be an available method for the separation and purification of phages.
Insights
A novel one-step salting-out extraction method efficiently purifies bacteriophages (phages) for treating multidrug-resistant bacterial infections. This cost-effective technique significantly improves phage recovery and impurity removal.
Area of Science:
- Microbiology
- Biotechnology
- Infectious Diseases
Background:
- Bacteriophages (phages) show promise as an alternative to antibiotics against multidrug-resistant bacteria.
- Traditional phage separation and purification methods are inefficient, costly, and time-consuming.
Purpose of the Study:
- To develop a simple, cost-saving, and efficient one-step salting-out extraction (SOE) method for purifying phage phiAB9.
- To optimize the SOE system for high phage recovery and impurity removal.
Main Methods:
- Investigated various salts and organic solvents for phage survival and SOE system compatibility.
- Optimized the SOE system composition using ammonium citrate and ethyl acetate.
- Evaluated phage recovery rate and removal rates of proteins, cells, and endotoxins.
Main Results:
- The optimized one-step SOE achieved a phage recovery rate of 90.82%.
- Achieved significant removal of impurities: 99.57% proteins, 97.98% cells, and 84.08% endotoxin.
- Demonstrated high concentration (210x) and purification factors for phage relative to impurities.
Conclusions:
- One-step salting-out extraction is an effective and efficient method for phage purification.
- This technique offers a viable alternative to traditional methods for preparing phages for therapeutic applications.
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