Background

As bacteriophages become increasingly important in therapeutic, agricultural, and biotechnological applications, the need for high purity, well-characterized phage preparations have never been more critical. Whether intended for clinical trials, commercial product development, or research use, phage preparations must meet stringent quality and safety standards. Impurities such as bacterial debris, endotoxins, host DNA, and media components not only compromise product efficacy, but can also induce adverse immune responses, limit regulatory approval, and reduce product shelf life.

Common phage purification techniques include clarification, precipitation, density gradient centrifugation, and dialysis. Key quality control methods comprise plaque assays, transmission electron microscopy (TEM), endotoxin testing, and SDS-PAGE.

At Creative BioMart Microbe, our phage purification and quality control services are designed to eliminate impurities such as bacterial debris, endotoxins, and host DNA, which could compromise phage efficacy and safety. We use advanced purification methods and rigorous quality control procedures to ensure that your phage preparations are safe, stable, and reproducible, making them suitable for clinical trials, commercial product development, and research. Contact us today to discuss your project needs or request a customized quote.

Service Procedure

Services Details

Our Advantages

• Regulatory Compliance: Our processes are designed to align with global regulatory expectations, including GMP and pharmacopeial standards.
• High Recovery Yields: Optimized protocols that maximize phage recovery while maintaining biological activity.
• Scalability: From milliliter-scale pilot studies to liter-scale production for preclinical or commercial use.
• Experienced Team: Expertise in handling a wide range of phage types, including dsDNA, RNA, lytic, and temperate phages.
• Custom Solutions: Tailored purification strategies based on your specific phage strain, application, and regulatory needs.
• Integrated Services: Seamless integration with our upstream services such as phage isolation, amplification, and formulation.

Case Study

Case Study 1: Purification of phage for therapeutic applications using high throughput anion exchange membrane chromatography.

With the rise of multidrug-resistant bacterial infections, bacteriophages are gaining renewed attention as an antibiotic alternative. However, current phage purification methods for therapeutic use remain inadequate, often requiring toxic reagents or costly equipment. In response, researchers have developed a rapid, scalable anion-exchange membrane chromatography method for purifying T7 phage from E. coli cultures. This one-step process eliminates the need for harmful chemicals and ultracentrifugation, achieving 65% phage recovery and up to 94% endotoxin removal in just 15 minutes. The method provides a safer, more efficient approach to phage purification and represents a significant advancement in clinical-grade phage production.

Figure 1. Schematic drawing showing strategy and working principles behind purification of bacteriophage using anion exchange membrane chromatography. (Roshankhah et al., 2023)

Figure 2. Phage recovery and endotoxin removal in experiments carried out by injecting 5 mL, 25 mL and 50 mL of 1:20 diluted feed solution. (Roshankhah et al., 2023)

Case Study 2: Generation of endotoxin-specific monoclonal antibodies by phage and yeast display for capturing endotoxin.

Using a phage display scFv library, monoclonal antibodies targeting E. coli O111:B4 LPS were selected by biopanning with biotinylated LPS on streptavidin-coated plates. High-affinity binders were identified by ELISA and sequencing, then reformatted into scFv-Fc and expressed in Expi293F cells. Two lead clones (P1 and P2) showed strong, specific LPS binding with minimal background as confirmed by SDS-PAGE and ELISA. This approach allows efficient isolation of LPS-specific antibodies for diagnostic or therapeutic use.

Figure 3. Generation of monoclonal antibodies targeting LPS from E. coli O111:B4 via phage display. (b) phage titer of panning rounds (1 to 3) for λ and κ sub libraries. (c) ELISA screening of 576 randomly picked individual clones. Clones that showed the lowest background signal (streptavidin/BSA) while displaying the highest LPS binding signal were chosen. Sequencing revealed recurrences. In orange: Clone P1, blue: Clone P2, and dark gray: Clone P3; dots in light gray represent excluded clones. (Fux, et al., 2024)

FAQs

References:

1. Fux AC, Casonato Melo C, Schlahsa L, et al. Generation of endotoxin-specific monoclonal antibodies by phage and yeast display for capturing endotoxin. IJMS. 2024;25(4):2297. doi:10.3390/ijms25042297
2. Roshankhah R, Jackson K, Nguyen TTN, Pelton R, Hosseinidoust Z, Ghosh R. Purification of phage for therapeutic applications using high throughput anion exchange membrane chromatography. Journal of Chromatography B. 2023;1229:123867. doi:10.1016/j.jchromb.2023.123867

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