Standard Bacteriophage Purification Procedures Cause Loss in Numbers and Activity

Carroll‐Portillo, Amanda; Coffman, Cristina; Varga, Matthew G.; Alcock, Joe; Singh, Sudha; Lin, Henry C.

doi:10.3390/v13020328

Cited by 47 publications

(40 citation statements)

References 44 publications

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“…Phages being used for commercial purposes need to be free of contaminants and impurities. These include, but not limited to bacteria, mold, lipopolysaccharides and host cell debris [73,74]. Before pre-formulation procedures, the crude lysate must be clarified and the phages captured, concentrated, and polished [72].…”

Section: Downstream Processing (Dsp)mentioning

confidence: 99%

“…Phages being used for commercial purposes need to b contaminants and impurities. These include, but not limited to bacteria, mold, l saccharides and host cell debris [73,74]. Before pre-formulation procedures, the c sate must be clarified and the phages captured, concentrated, and polished [72 steps constitute the major unit operations for phages as they relate to biologics m turing and serve to isolate and concentrate the phages for further processing [72 overview schematic refer to Figure 5.…”

Section: Downstream Processing (Dsp)mentioning

confidence: 99%

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Manufacturing Bacteriophages (Part 1 of 2): Cell Line Development, Upstream, and Downstream Considerations

et al. 2021

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Within this first part of the two-part series on phage manufacturing, we will give an overview of the process leading to bacteriophages as a drug substance, before covering the formulation into a drug product in the second part. The principal goal is to provide the reader with a comprehensive framework of the challenges and opportunities that present themselves when developing manufacturing processes for bacteriophage-based products. We will examine cell line development for manufacture, upstream and downstream processes, while also covering the additional opportunities that engineered bacteriophages present.

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Section: Downstream Processing (Dsp)mentioning

confidence: 99%

Section: Downstream Processing (Dsp)mentioning

confidence: 99%

Manufacturing Bacteriophages (Part 1 of 2): Cell Line Development, Upstream, and Downstream Considerations

et al. 2021

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“…A common operating procedure for varied phage preparations, storage, and transport is lacking [51]. Standard operating procedure (s) for large scale-bacteriophage cultivation, isolation, titration, and purification and to produce sufficient plaqueforming units of bacteriophages (PFUs) per milliliter of Pseudomonas, Klebsiella, and Serratia were established [41,52]. Such a universal process and production of the final phage preparations for use could reduce endotoxins, might be pivotal in alleviating fears and the phage therapy shall be readily accepted all the world over.…”

Section: Therapeutic Bacteriophagesmentioning

confidence: 99%

“…Phage (VPP97) of V. parahaemolyticus have been shown to be stable up to 65°C and were totally inactivated at 70°C [10,[61][62][63][64]. Bacteriophages were detected to survive extremes over 95°C [52]. Bacteriophages such as T-ϕD0, T-ϕD2S, T-ϕHSIC, and T-ϕD1B exhibited a latent period ranging beyond 90°C [64].…”

Section: Storage Of Phagesmentioning

confidence: 99%

Therapeutic Efficacy of Bacteriophages

Ramasamy¹,

Dayanithi²

2021

Bacteriophages in Therapeutics

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Bacteriophages are bacterial cell-borne viruses that act as natural bacteria killers and they have been identified as therapeutic antibacterial agents. Bacteriophage therapy is a bacterial disease medication that is given to humans after a diagnosis of the disease to prevent and manage a number of bacterial infections. The ability of phage to invade and destroy their target bacterial host cells determines the efficacy of bacteriophage therapy. Bacteriophage therapy, which can be specific or nonspecific and can include a single phage or a cocktail of phages, is a safe treatment choice for antibiotic-resistant and recurrent bacterial infections after antibiotics have failed. A therapy is a cure for health problems, which is administered after the diagnosis of the diseases in the patient. Such non-antibiotic treatment approaches for drug-resistant bacteria are thought to be a promising new alternative to antibiotic therapy and vaccination. The occurrence, biology, morphology, infectivity, lysogenic and lytic behaviours, efficacy, and mechanisms of bacteriophages’ therapeutic potentials for control and treatment of multidrug-resistant/sensitive bacterial infections are discussed. Isolation, long-term storage and recovery of lytic bacteriophages, bioassays, in vivo and in vitro experiments, and bacteriophage therapy validation are all identified. Holins, endolysins, ectolysins, and bacteriocins are bacteriophage antibacterial enzymes that are specific. Endolysins cause the target bacterium to lyse instantly, and hence their therapeutic potential has been explored in “Endolysin therapy.” Endolysins have a high degree of biochemical variability, with certain lysins having a wider bactericidal function than antibiotics, while their bactericidal activities are far narrower. Bacteriophage recombinant lysins (chimeric streptococcal–staphylococcal constructs) have high specificity for a single bacterial species, killing only that species (lysin (CF-301) is focused to kill methicillin resistant Staphylococcus aureus (MRSA)), while other lysins have a broader lytic activity, killing several different bacterial species and hence the range of bactericidal activity. New advances in medicine, food safety, agriculture, and biotechnology demonstrate molecular engineering, such as the optimization of endolysins for particular applications. Small molecule antibiotics are replaced by lysins. The chapter discusses the occurrences of lytic phage in pathogenic bacteria in animals and humans, as well as the possible therapeutic effects of endolysins-bacteriophage therapy in vivo and in vitro, demonstrating the utility and efficacy of the therapy. Further developments in the bacteriophage assay, unique molecular-phage therapy, or a cocktail of phage for the control of a broad range of drug-resistant bacteria-host systems can promote non-antibiotic treatment methods as a viable alternative to conventional antibiotic therapy.

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“…Additionally, as phages are highly sensitive to shear stress, the mixing process must take that into account. Furthermore, the viscosity of the formula can also be a concern as highly viscous fluids tend to offer higher shear stress, which can cause phage inactivation [ 2 , 8 , 9 , 18 , 33 , 34 ].…”

Section: Introductionmentioning

confidence: 99%

Manufacturing Bacteriophages (Part 2 of 2): Formulation, Analytics and Quality Control Considerations

et al. 2021

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Within this second piece of the two-part series of phage manufacturing considerations, we are examining the creation of a drug product from a drug substance in the form of formulation, through to fill-finish. Formulation of a drug product, in the case of bacteriophage products, is often considered only after many choices have been made in the development and manufacture of a drug substance, increasing the final product development timeline and difficulty of achieving necessary performance parameters. As with the preceding review in this sequence, we aim to provide the reader with a framework to be able to consider pharmaceutical development choices for the formulation of a bacteriophage-based drug product. The intent is to sensitize and highlight the tradeoffs that are necessary in the development of a finished drug product, and to be able to take the entire spectrum of tradeoffs into account, starting with early-stage R&D efforts. Furthermore, we are arming the reader with an overview of historical and current analytical methods with a special emphasis on most relevant and most widely available methods. Bacteriophages pose some challenges that are related to but also separate from eukaryotic viruses. Last, but not least, we close this two-part series by briefly discussing quality control (QC) aspects of a bacteriophage-based product, taking into consideration the opportunities and challenges that engineered bacteriophages uniquely present and offer.

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Standard Bacteriophage Purification Procedures Cause Loss in Numbers and Activity

Cited by 47 publications

References 44 publications

Manufacturing Bacteriophages (Part 1 of 2): Cell Line Development, Upstream, and Downstream Considerations

Manufacturing Bacteriophages (Part 1 of 2): Cell Line Development, Upstream, and Downstream Considerations

Therapeutic Efficacy of Bacteriophages

Manufacturing Bacteriophages (Part 2 of 2): Formulation, Analytics and Quality Control Considerations

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