Background

Research-grade phage production is a critical process for advancing phage therapy, diagnostics, and basic microbiological research. As phage therapy progresses towards clinical applications, the need for high-quality, well-characterized, and scalable phage production methods has become increasingly important. This involves ensuring that phages are free from contaminants, antibiotic resistance genes, and other impurities, while maintaining their efficacy and stability. Key steps in phage production include:

Upstream Processing (USP)

• Cell Line Development: The first step involves creating a master cell bank and a master phage bank. This ensures consistency and reproducibility in phage production.
• Production Modes: Phage production can be carried out in batch, semi-continuous, or continuous modes. Batch processes are cost-effective and can achieve high titers but may suffer from variability and high downtime. Semi-continuous processes combine the advantages of batch and continuous systems, improving efficiency and reducing variability.
• Single-Use vs. Stainless Steel Systems: Single-use technologies (SUT) are increasingly popular due to their ability to reduce cross-contamination, decrease cleaning and validation steps, and increase overall productivity.

Downstream Processing (DSP)

• Purification: After phage amplification, downstream processing is crucial for removing contaminants and achieving pure phage preparations. Techniques such as precipitation with polyethylene glycol (PEG) are commonly used to concentrate phages.

Formulation and Storage

• Solid Formulations: Solid formulations such as pills, capsules, and coated devices are preferred for patient convenience. Techniques like lyophilization and spray-drying are used to stabilize phage preparations.
• Encapsulation for Targeted Delivery: Encapsulation in alginate or liposomes can protect phages during transit through the gastrointestinal tract and improve their residence time at the target site.

Figure 1. Overview of phage manufacturing. (João et al., 2021)

At Creative BioMart Microbe, our research-grade phage production services are designed to support scientists and innovators in obtaining high-quality, customizable phage preparations tailored to their specific research needs. Whether you're exploring phage therapy, developing diagnostic tools, or studying bacterial dynamics, our services provide the foundation for reliable and reproducible results. For more information or to request a quote, please contact us.

Service Procedure

Services Details

Custom Phage Production

We offer tailored phage production services to meet your specific research needs. Our team works closely with you to understand your requirements and design a production plan that ensures high-quality phage preparations. From small-scale laboratory needs to larger quantities for extensive studies, our flexible production capabilities and stringent quality controls guarantee consistent results.

High-Purity Phage Isolation

Our advanced purification protocols ensure that the phages we produce are of the highest purity. Utilizing state-of-the-art techniques such as ultracentrifugation, chromatography, and tangential flow filtration, we remove contaminants and concentrate phage particles effectively. Each batch undergoes rigorous testing to confirm purity levels, ensuring that your research is supported with reliable and consistent phage preparations.

Comprehensive Quality Control

Quality is at the core of our services. Every step of the phage production process is monitored and validated through comprehensive quality control measures. Our team employs a range of analytical instruments, including TEM, AFM, NGS, and MS, to characterize and verify the phages. Stringent QC protocols ensure that each batch meets stringent standards for purity, potency, and consistency, providing you with reliable phage products for your research.

Scalable Solutions

Whether you need small quantities for initial studies or larger volumes for extensive research, our scalable production platforms can accommodate your needs. Our flexible bioreactor systems and disposable fermentation solutions allow for seamless scale-up without compromising quality. We work with you to develop a scalable production plan, ensuring that your phage supply can grow with your research demands.

Platforms and Instrumentation

Our Advantages

• Customized Solutions: Tailored phage production to match your specific research requirements and objectives.
• High Purity Standards: Rigorous purification protocols ensure minimal contaminants, suitable for sensitive applications.
• Scalable Production: Flexible batch sizes accommodate both small-scale studies and larger research projects.
• Expertise in Phage Biology: Our team possesses extensive experience in phage research, ensuring informed and effective production strategies.
• Rapid Turnaround: Efficient processes and streamlined workflows enable timely delivery of phage preparations.
• Comprehensive Quality Control: Each batch undergoes thorough testing to guarantee consistency and reliability.

Case Study

Case Study 1: Two-stage self-cycling (TSSC) process for phage amplification in bioreactors.

To combat Cronobacter sakazakii, a deadly infant pathogen increasingly resistant to antibiotics, researchers developed a high-yield phage production method using a two-stage self-cycling (TSSC) process. By monitoring pH in real time, they pinpointed optimal conditions: introduce phages when pH hits 5.8 (early log phase) and harvest at 4.94. This streamlined process allowed phages to multiply efficiently without repeated manual intervention. Compared to traditional batch culture, the TSSC method produced 24× higher phage titers using the same volume of LB medium and maintained stable yields across three production cycles.

Figure 2. Optimal initial infection conditions for phage Φ CS01 production. (Lee et al., 2021)

Case Study 2: Erwinia amylovora phage fermentation optimization.

This study tackled the challenge of industrial-scale phage production for fighting Erwinia amylovora, the culprit behind fire blight—especially important given rising antibiotic resistance. Researchers focused on optimizing seed culture to improve safety and scalability. By fine-tuning inoculum levels and supplementing media with fructose and sucrose, they achieved phage yields comparable to traditional methods but with significantly lower inputs (just 10⁵ CFU/mL bacteria and 10³ PFU/mL phages). Further pH adjustments in the fermenter boosted yields by up to 303%.

Researchers tested the effect of different initial phage concentrations (inoculum) on the production yield of three model phages: pEa_8, pEa_27, and pEa_31. Lower inoculum levels led to significantly higher yields for all three phages. pEa_8 saw yield increases of up to 999.66% as the inoculum decreased, pEa_27 achieved over 200% yield improvement at 3–5 Log PFU/mL, and pEa_31 peaked at a 1,206.99% yield increase at 3 Log PFU/mL. All phages performed best at lower inoculum concentrations. The highest production was achieved at 3 Log PFU/mL. This concentration was selected for future experiments.

Figure 3. Phage production yields for Erwinia amylovora phages at different inoculum concentrations. Yields for the following E. amylovora phages were measured: pEa_SNUABM_8 (a), pEa_SNUABM_27 (b), and pEa_SNUABM_31 (c). (Jo et al., 2024)

FAQs

References:

1. Jo SJ, Giri SS, Lee SB, et al. Optimization of the large-scale production for Erwinia amylovora bacteriophages. Microb Cell Fact. 2024;23(1):342.
2. João J, Lampreia J, Prazeres DMF, Azevedo AM. Manufacturing of bacteriophages for therapeutic applications. Biotechnology Advances. 2021;49:107758.
3. Lee JS, Kim GH, Kim J, Lim TH, Yoon Y, Yoon SS. Large-scale production of Cronobacter sakazakii bacteriophage φ CS01 in bioreactors via a two-stage self-cycling process. J Microbiol Biotechnol. 2021;31(10):1430-1437.