Background

Phage production services include the processes involved in the development, amplification, purification, characterization and formulation of bacteriophages. Although bacteriophages have been studied since the early 20th century, their therapeutic and biotechnological potential has received renewed attention in recent decades, primarily due to the escalating global challenge of antimicrobial resistance (AMR) and the increasing demand for targeted, sustainable biocontrol strategies in various sectors, including healthcare, agriculture, and food safety.

Comprehensive phage production services typically include upstream and downstream processing. Among them, there are different production modes.

Upstream Processing (USP)

The transition from laboratory to industrial scale production involves several critical steps:

• Inoculum Preparation: Cultivation of host bacteria and preparation of phage inoculum under controlled conditions.
• Fermentation: Infecting the host bacteria with phages in bioreactors, optimizing parameters such as temperature, pH, and oxygen levels to maximize phage yield.
• Monitoring and Control: Implementation of real-time monitoring systems to ensure that the process remains within defined parameters, in line with the desired Quality Target Product Profile (QTPP).

Production Modes

Choosing the right production mode is critical to optimizing yield and process efficiency:

• Batch Process: All components are combined in a single run and the product is harvested at the end. This method is straightforward, but may have scalability limitations.
• Semi-Continuous Process: Involves partial harvesting and replenishment of the culture, allowing for extended production periods and improved resource utilization.
• Continuous Process: A steady-state operation in which fresh media is continuously added while culture is simultaneously removed. This mode provides consistent product quality and is suitable for large-scale production.

Figure 1. Production modes for phage manufacturing: batch, semi-continuous, and fully continuous. (Adapted from Tanir et al., 2021)

Downstream Processing (DSP)

After production, the phage-containing broth is purified to meet therapeutic standards:

• Clarification: Removal of bacterial cells and debris by centrifugation or filtration.
• Concentration: Using techniques such as ultrafiltration to concentrate the phage particles.
• Purification: Removal of contaminants such as endotoxins using chromatography or other purification methods.
• Formulation: Stabilization of purified phages in appropriate buffers for storage and administration.

From isolation to industrial-scale production, Creative BioMart Microbe offers a full range of phage production services tailored to your application, whether you're developing next-generation personalized phage therapies, pioneering biocontrol strategies in food and agriculture, or fine-tuning microbiome modulation. Our state-of-the-art capabilities ensure consistency, purity, and scalability at every stage of production. Contact us today to streamline your production pipeline with expert support and industry-grade quality.

Service Procedure

Service Details

Our Advantages

• Phage Expertise: Over a decade of experience with diverse phage families and host systems in both research and industrial contexts.
• Flexible Scaling: Fermentation capacity from R&D batches (1 L) to industrial volumes (100 L+), with fast turnaround.
• Customized Processes: Tailored protocols for unique phage-host systems, including anaerobic fermentation for gut-specific phages.
• Integrated Services: Complete pipeline from isolation to formulation, reducing handoffs and accelerating time to product.

Case Study

Case Study 1: Cell-free production of personalized therapeutic phages targeting multidrug-resistant bacteria.

This study presents a comprehensive cell-free platform for rapid, personalized production, transient bioengineering, and proteomic analysis of diverse phages. Using mass spectrometry, the researchers validated previously hypothesized and non-structural proteins and monitored phage protein expression during assembly. Remarkably, small-volume reactions yielded therapeutically effective phages targeting Escherichia coli (EAEC), Yersinia pestis, and Klebsiella pneumoniae. The versatility of the platform was further demonstrated by adapting it to produce phages against Gram-positive bacteria by co-expression of specific host factors. In addition, a novel non-genomic engineering approach was introduced to enable single cycle functional modifications. Overall, this method provides a promising foundation for both reverse and forward phage engineering and the custom production of clinical-grade phage therapies.

Figure 2. Schematic of 'phactory', a cell-free platform for reverse and forward engineering of therapeutic phages. (Emslander et al., 2022)

Case Study 2:Strategy for mass production of lytic Staphylococcus aureus bacteriophage pSa-3.

This study focused on optimizing the production of Staphylococcus aureus phage pSa-3 by manipulating host and phage inoculum concentrations and energy sources to extend the co-culture phase and enhance yield. By reducing the phage multiplicity of infection (MOI) and adjusting bacterial inoculum levels, researchers delayed bacterial lysis and identified optimal conditions—2 × 10⁸ CFU/mL bacterial inoculum and 0.0001 MOI—achieving a maximum phage yield of 1.68 × 10¹⁰ PFU/mL. Further optimization using response surface methodology, with glycerol, glycine, and calcium as selected nutrient sources, resulted in a final concentration of 8.63 × 10¹⁰ ± 9.71 × 10⁹ PFU/mL—a 5.13-fold increase. These findings demonstrate that adjusting inoculum ratios and employing statistical optimization tools can significantly improve the efficiency and scalability of lytic bacteriophage production.

Figure 3. Growth curve of the microorganisms in the bacteria–phage co-culture. Bacterial growth was measured using optical density (a, c, e). Phage growth was measured in PFU count (b, d, f). 2 × 10⁸ CFU/mL of initial Staphylococcus aureus concentration (a, b), 1 × 10⁹ CFU/mL of S. aureus concentration (c, d), and 2 × 10⁹ CFU/mL of S. aureus concentration (e, f). (Kim et al., 2021)

FAQs

References:

1. Emslander Q, Vogele K, Braun P, et al. Cell-free production of personalized therapeutic phages targeting multidrug-resistant bacteria. Cell Chemical Biology. 2022;29(9):1434-1445.e7. doi:10.1016/j.chembiol.2022.06.003
2. Kim SG, Kwon J, Giri SS, et al. Strategy for mass production of lytic Staphylococcus aureus bacteriophage pSa-3: contribution of multiplicity of infection and response surface methodology. Microb Cell Fact. 2021;20(1):56. doi:10.1186/s12934-021-01549-8
3. Tanir T, Orellana M, Escalante A, Moraes De Souza C, Koeris MS. Manufacturing bacteriophages (Part 1 of 2): cell line development, upstream, and downstream considerations. Pharmaceuticals. 2021;14(9):934. doi:10.3390/ph14090934

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