Optimizing phage amplification
The issue of low PFU/ml results when amplifying phages is a common problem facing researchers worldwide. With a focus on quality over quantity at the early development stage, this is often found to be an issue when scaling up for product trials and for full scale manufacturing. Increasing PFU/ml at this stage provides more phage product for use in trials or for selling to clients.
Using flasks for phage amplification
At the research level, amplification of phages in laboratories is commonly carried out in Erlenmeyer flasks, with air being pumped into the media using a pipette as shown in the diagram below. The flask is then transferred to an incubator for temperature control.
While this technique produces phage product, there are inherent drawbacks to the method which contribute to low and inconsistent PFU/ml results.
Difficulties of working with flasks
- Lack of control due to empiric nature of the flask
- Inability to control temperature effectively in the incubator
- Inability to control gas flow at optimum rate
- Risk of contamination
- Significant downtime between each experiment with cleaning and sterilization required
- Difficult to scale up
Important parameters to control
Working with client trials, we discovered that the following parameters are crucial in optimizing phage amplification:
- Temperature control
- Gas Flow rate
- OD measurements
- Contamination reduction
Using the CellMaker to optimize phage amplification
Having identified these issues and key parameters, we have developed a new solution to improve phage manufacturing. The CellMaker uses a patented, revolutionary airlift technology which provides optimal aeration, and our single-use technology reduces downtime and eliminates the need for cleaning.
The CellMaker has several key benefits to optimize phage amplification:
- Accurate temperature control to +/- 0.1C provided by a Peltier plate situated within the enclosure unit
- Gas flow rate measured by mass flow controllers, with a tolerance of 0.005L/min, providing the accuracy required for phage amplification
- Real-time optical density measurements from in-situ sensors
- Reduction in contamination due to sterile bioreactors fitted with tubing and connections for “plug and play” convenience
- Reduced downtime due to single use technology, eliminating the need for cleaning and sterilization
- Flexible solution, with working volumes from 3L to 50L
- Further downstream processing can be achieved; a discussion with Cellexus would facilitate this
- Ready to use in a GMP facility with CFR chapter 11-compliant software
Results from the CellMaker
Our technology is flexible and can be used across industries. The CellMaker is currently in use in the animal health, food safety, crop protection and drug discovery industries. Our technology has successfully amplified phages from E. coli, Salmonella, Listeria, Pseudomonas aeruginosa, Pseudomonas syringae, Staphylococcus aureus and Yersinia ruckeri, all producing exciting results. A selection of these results are seen below:
||5 × 107
||1 × 1010
||1 × 105
||1 × 108
||1 × 109
||3 × 1010
||1 x 109
||3 × 1010
Reduced manufacturing costs
Manufacturing cost is key to the success or failure of a product, and the significance of choosing the best technology for scaling up your process cannot be underestimated.
The CellMaker is producing exciting phage amplification results due to the ability to control process parameters, the speed and ease of single-use technology, and the efficiency of the airlift process. This results in optimized PFU/ml results and a faster route to market.
Read more about our results in our case studies.