High-Purity Phage Preparation Services
BackgroundService ProcedureOur AdvantagesCase StudyFAQs
Background
High-purity phage preparation involves isolating bacteriophages to a level of purity suitable for therapeutic, research, and industrial applications. Achieving this is vital for ensuring safety, efficacy, and experimental accuracy. Traditionally, phage samples were contaminated with bacterial residues, such as endotoxins, due to limitations in basic filtration and centrifugation techniques. These traditional methods often fell short of the purity levels required for clinical or pharmaceutical use.
Today, modern purification techniques have significantly advanced. The double agar layer method allows cleaner plaque isolation, while membrane filtration and high-speed centrifugation effectively remove bacterial contaminants. Chemical treatments, including lysozyme and chelating agents, further degrade residual cell material. To verify purity, methods like SDS-PAGE, HPLC, and endotoxin testing (e.g., LAL assay) are employed, ensuring that phage preparations meet rigorous quality standards for advanced applications.
Figure 1. Flow chart of the Phage On Tap protocol for the production of high titer, homogenous, endotoxin reduced phage banks within two days. (Bonilla et al., 2016)
Applications
High-purity phage preparations have a wide range of applications, including:
- Phage Therapy: Treating bacterial infections, especially those resistant to antibiotics.
- Research: Studying phage biology, genetics, and interactions with bacteria.
- Industrial Use: In biotechnology and pharmaceutical industries for the development of new therapies and products.
Creative BioMart Microbe offers high-purity phage preparation services, providing optimized and scalable workflows for delivering phages of exceptional quality. Our robust platform integrates classical virology techniques and modern purification technologies. We are committed to producing phage preparations that meet the stringent demands of translational research and product development. Contact us today for more information!
Service Procedure
Service Details
| Service |
Type |
Description |
| Optimized Phage Amplification |
Scalable Phage Propagation |
Propagation in selected host strains under tightly controlled conditions. |
| MOI and Culture Optimization |
Optimization of multiplicity of infection (MOI), culture density, and incubation for maximum yield. |
| Lytic and Engineered Phage Support |
Applicable for both naturally lytic and genetically engineered phages. |
| Multi-Step Purification Workflow |
Centrifugation and Filtration |
Removal of cell debris and bulk host contaminants. |
| PEG Precipitation or TFF |
Phage concentration and buffer exchange. |
| CsCl or Iodixanol Gradient Ultracentrifugation (optional) |
High-resolution separation of phage particles from proteins, DNA, and endotoxins. |
| Endotoxin Remove |
Critical for phage preparations used in therapeutic and in vivo applications. |
| Analytical Characterization |
Titer Determination |
Quantification using spot assays or double-layer plaque methods (PFU/mL). |
| Purity Assessment |
Analysis via SDS-PAGE, OD260/280 ratio, and optional TEM imaging. |
| Endotoxin Testing |
LAL assay or equivalent methods, with customizable thresholds. |
| Sterility Testing |
Ensures absence of host bacteria, fungi, or mycoplasma contamination. |
Our Advantages
- Endotoxin-Free Formulations: Achieve endotoxin levels below 0.1 EU/mL, meeting the requirements for preclinical and translational studies.
- Customizable Purification Protocols: We offer density gradients, chromatography, and endotoxin reduction tailored to the physical and biochemical properties of each phage.
- Minimal Host DNA Contamination: Host nucleic acid levels are minimized through nuclease treatments and confirmed by sensitive DNA quantification methods.
- Flexible and Scalable Workflows: Our processes are adaptable for small-scale research as well as pilot-scale manufacturing preparations.
- Comprehensive QC Testing: Each batch is accompanied by a full set of quality control data, ensuring transparency, traceability, and reproducibility.
- Rapid Turnaround: Our optimized pipeline enables delivery within 2–3 weeks, depending on project complexity, without compromising quality.
Case Study
Case Study 1: Chronic bacterial prostatitis treated with phage therapy after multiple failed antibiotic treatments.
A patient with chronic bacterial prostatitis (CBP) failed multiple antibiotic treatments due to infection with multidrug-resistant pathogens, including MRSA, Enterococcus faecalis, and Streptococcus mitis. Phage therapy, using high-purity bacteriophage preparations from the Eliava Institute, was administered following phage sensitivity testing. The patient experienced significant symptom relief and a marked reduction in bacterial load. This case underscores the potential of purified phage therapy in treating persistent, biofilm-associated infections in anatomically challenging, low-vascularity sites—especially when antibiotic options are limited.
Table 1. Standard phage preparations made by the Eliava institute of bacteriophages, microbiology and virology. (Johri et al., 2021)
Case Study 2: Design of phage-cocktail–containing hydrogel for the treatment of Pseudomonas aeruginosa–infected wounds.
Due to rising antibiotic resistance, treating Pseudomonas aeruginosa wound infections is increasingly challenging. This study developed a phage-loaded hydrogel dressing using two polyvalent phages. In mouse models, phage hydrogels showed antimicrobial effects comparable to antibiotics and promoted better wound healing. The combination of phages and antibiotics had the strongest effect, highlighting phage hydrogels as a promising alternative for managing antibiotic-resistant wound infections.
Figure 1. Plaques of phages (A) PB10 (107 PFU/mL) and (B) PA19 (107 PFU/mL) on P. aeruginosa ATCC27853. (Shafigh Kheljan et al., 2023)
FAQs
Q: What types of phages do you support?
A: We routinely work with lytic phages targeting both Gram-positive and Gram-negative bacteria. Please contact us regarding unusual or temperate phages.
Q: What is the minimum starting volume required?
A: We recommend at least 10–100 mL of lysate, though we can work with lower volumes for pilot studies or rare phages.
Q: What level of purity do your phage preparations achieve?
A: Our phage preparations undergo multi-step purification processes, including tangential flow filtration (TFF), nuclease treatment, and endotoxin removal, achieving high purity with minimal residual host DNA, proteins, and endotoxins—meeting standards suitable for preclinical and clinical applications.
Q: Can you tailor purification protocols to our phage and application?
A: Yes, we customize purification workflows based on your phage type, host strain, and intended use—whether for in vivo studies, therapeutic development, or regulatory submission—to balance yield, purity, and biological activity.
Q: What types of analytical testing do you offer with purified phage products?
A: We offer a full suite of QC assays, including endotoxin levels (LAL test), residual host DNA/protein quantification, sterility testing, titer accuracy, and electron microscopy upon request.
Q: What is the typical turnaround time?
A: Standard processing time is approximately 2–3 weeks. Timelines may vary based on phage complexity and purification method.
Q: Do you accommodate confidentiality agreements?
A: Yes. We are happy to sign NDAs or provide standard confidentiality agreements as part of the onboarding process.
References:
- Bonilla N, Rojas MI, Netto Flores Cruz G, Hung SH, Rohwer F, Barr JJ. Phage on tap–a quick and efficient protocol for the preparation of bacteriophage laboratory stocks. PeerJ. 2016;4:e2261. doi:10.7717/peerj.2261
- Johri AV, Johri P, Hoyle N, Pipia L, Nadareishvili L, Nizharadze D. Case report: chronic bacterial prostatitis treated with phage therapy after multiple failed antibiotic treatments. Front Pharmacol. 2021;12:692614. doi:10.3389/fphar.2021.692614
- Shafigh Kheljan F, Sheikhzadeh Hesari F, Aminifazl M, Skurnik M, Goladze S, Zarrini G. Design of phage-cocktail–containing hydrogel for the treatment of Pseudomonas aeruginosa–infected wounds. Viruses. 2023;15(3):803. doi:10.3390/v15030803
