Effect of dilution rate on productivity of continuous bacteriophage production in cellstat

Nabergoj, Dominik; Kuzmić, Nina; Drakslar, Benjamin; Podgornik, Aleš

doi:10.1007/s00253-018-8893-9

Cited by 19 publications

(18 citation statements)

References 62 publications

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“…The downsides for industrial scale batch fermentation include higher capital costs, large process footprints, labour-intensive operation, that the proportion of downtime compared with production time can be high, a lack of process control, and variability of product quality [ 24 ]. The continuous upstream production of phages using chemostat systems has heretofore received little attention in the published literature, which instead has focused on using such systems for studying coevolution processes [ 25 , 26 , 27 ]. Decoupling the bacterial host propagation from phage production removes the selection pressure for bacteria mutation and should allow stable long-term steady-state operation of the process [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

High Throughput Manufacturing of Bacteriophages Using Continuous Stirred Tank Bioreactors Connected in Series to Ensure Optimum Host Bacteria Physiology for Phage Production

Mancuso

Shi

Malik

2018

Viruses

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Future industrial demand for large quantities of bacteriophages e.g., for phage therapy, necessitates the development of scalable Good Manufacturing Practice compliant (cGMP) production platforms. The continuous production of high titres of E coli T3 phages (1011 PFU mL−1) was achieved using two continuous stirred tank bioreactors connected in series, and a third bioreactor was used as a final holding tank operated in semi-batch mode to finish the infection process. The first bioreactor allowed the steady-state propagation of host bacteria using a fully synthetic medium with glucose as the limiting substrate. Host bacterial growth was decoupled from the phage production reactor downstream of it to suppress the production of phage-resistant mutants, thereby allowing stable operation over a period of several days. The novelty of this process is that the manipulation of the host reactor dilution rates (range 0.1–0.6 hr−1) allows control over the physiological state of the bacterial population. This results in bacteria with considerably higher intracellular phage production capability whilst operating at high dilution rates yielding significantly higher overall phage process productivity. Using a pilot-scale chemostat system allowed optimisation of the upstream phage amplification conditions conducive for high intracellular phage production in the host bacteria. The effect of the host reactor dilution rates on the phage burst size, lag time, and adsorption rate were evaluated. The host bacterium physiology was found to influence phage burst size, thereby affecting the productivity of the overall process. Mathematical modelling of the dynamics of the process allowed parameter sensitivity evaluation and provided valuable insights into the factors affecting the phage production process. The approach presented here may be used at an industrial scale to significantly improve process control, increase productivity via process intensification, and reduce process manufacturing costs through process footprint reduction.

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Section: Introductionmentioning

confidence: 99%

High Throughput Manufacturing of Bacteriophages Using Continuous Stirred Tank Bioreactors Connected in Series to Ensure Optimum Host Bacteria Physiology for Phage Production

Mancuso

Shi

Malik

2018

Viruses

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“…Here, we will analyze first theoretical studies focused on phage production models and then selected studies that have been validated experimentally. All the cases agree with the assay criteria for further purification and validation of a bacteriophage-based product, and some of them are included in both sections (Santos et al, 2014; Nabergoj et al, 2018a).…”

Section: Generalities In Bacteriophage Productionmentioning

confidence: 60%

“…This level of production agrees with the production needed for therapeutic purposes (>1 10 10 PFU mL –1 ), considering purification steps, the decay rate of phages and stability or shelf life (Naghizadeh et al, 2018). Other authors have also reported promising levels of production of 5 × 10 12 PFU mL –1 in 1.2 L (Warner et al, 2014), and 2.4 × 10 13 PFU day –1 in 1 L (Nabergoj et al, 2018a; Table 2).…”

Section: Experimental Experiences In Bacteriophage Productionmentioning

confidence: 87%

“…Besides, regulating the dilution rate will allow direct control over the bacterial growth rate, which has a direct influence on infection parameters like burst size, adsorption constant and latent period (Mancuso et al, 2018; Nabergoj et al, 2018b). Dilution rate can also be used to increase the productivity of the system, as was shown by Nabergoj et al (2018a) where a maximum phage productivity of 10 9 phages mL –1 h –1 was achieved with a low dilution rate of 2 h –1 in a 1 L cellstat system. A continuous operating system can be permanently operative, and is therefore the most convenient way to produce a biotechnological product for a company.…”

Section: Experimental Experiences In Bacteriophage Productionmentioning

confidence: 99%

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Bacteriophage Production Models: An Overview

et al. 2019

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The use of bacteriophages has been proposed as an alternative method to control pathogenic bacteria. During recent years several reports have been published about the successful use of bacteriophages in different fields such as food safety, agriculture, aquaculture, and even human health. Several companies are now commercializing bacteriophages or bacteriophage-based products for therapeutic purposes. However, this technology is still in development and there are challenges to overcome before bacteriophages can be widely used to control pathogenic bacteria. One big hurdle is the development of efficient methods for bacteriophage production. To date, several models for bacteriophage production have been reported, some of them evaluated experimentally. This mini-review offers an overview of different models and methods for bacteriophage production, contrasting their principal differences.

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“…Although chemostat allows the cultivation of microorganisms at a physiological steady state [ 31 ], the bacterial culture may be less genetically stable, as mutations can occur [ 32 ]. Cellstat is recognised as a phage production system for strictly lytic phages [ 33 ] that avoids direct phage exposure and pressure but requires the use of two different bioreactors [ 34 ].…”

Section: Introductionmentioning

confidence: 99%

Production of Bacteriophages by Listeria Cells Entrapped in Organic Polymers

Roy

Philippe

Loessner

et al. 2018

Viruses

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Applications for bacteriophages as antimicrobial agents are increasing. The industrial use of these bacterial viruses requires the production of large amounts of suitable strictly lytic phages, particularly for food and agricultural applications. This work describes a new approach for phage production. Phages H387 (Siphoviridae) and A511 (Myoviridae) were propagated separately using Listeria ivanovii host cells immobilised in alginate beads. The same batch of alginate beads could be used for four successive and efficient phage productions. This technique enables the production of large volumes of high-titer phage lysates in continuous or semi-continuous (fed-batch) cultures.

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Effect of dilution rate on productivity of continuous bacteriophage production in cellstat

Cited by 19 publications

References 62 publications

High Throughput Manufacturing of Bacteriophages Using Continuous Stirred Tank Bioreactors Connected in Series to Ensure Optimum Host Bacteria Physiology for Phage Production

High Throughput Manufacturing of Bacteriophages Using Continuous Stirred Tank Bioreactors Connected in Series to Ensure Optimum Host Bacteria Physiology for Phage Production

Bacteriophage Production Models: An Overview

Production of Bacteriophages by Listeria Cells Entrapped in Organic Polymers

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