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.
复制标题
DOI：
10.3390/v10100537
复制
发表时间：
2018-10-01
期刊：
影响因子：
--
通讯作者：
Malik DJ
中科院分区：
文献类型：
--
作者：
Mancuso F;Shi J;Malik DJ
未来工业对大量噬菌体的需求，例如用于噬菌体治疗，需要开发符合良好制造规范(CGMP)的可扩展生产平台。利用串联的两个连续搅拌槽生物反应器和第三个生物反应器作为半间歇操作的终止罐，实现了大肠杆菌T3噬菌体高滴度(1011pfu m L−1)的连续生产。第一个生物反应器允许宿主细菌使用以葡萄糖为限制底物的全合成介质进行稳定繁殖。宿主细菌的生长与其下游的噬菌体生产反应器脱钩，以抑制抗噬菌体突变体的产生，从而允许在几天内稳定运行。这一过程的新奇之处在于，通过操纵宿主反应器的稀释率(范围为0.1hr−1)，可以控制细菌种群的生理状态。这导致细菌具有相当高的胞内噬菌体生产能力，同时在高稀释率下操作，产生显着更高的总体噬菌体过程生产率。使用中试规模的恒化器系统可以优化上游噬菌体扩增条件，有利于宿主细菌的高细胞内噬菌体产量。考察了宿主反应器稀释率对噬菌体爆发量、滞留时间和吸附速率的影响。寄主细菌的生理特性影响噬菌体的大小，从而影响整个过程的生产率。对过程的动力学进行数学建模，可以对参数敏感性进行评估，并为影响噬菌体生产过程的因素提供有价值的见解。这里提出的方法可以在工业规模上使用，以显著改善过程控制，通过过程强化提高生产率，并通过减少过程足迹来降低过程制造成本。
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.
登录
查看更多内容
影响因子：
46.9
作者：
Bikard, David;Euler, Chad W.;Jiang, Wenyan;Nussenzweig, Philip M.;Goldberg, Gregory W.;Duportet, Xavier;Fischetti, Vincent A.;Marraffini, Luciano A.
通讯作者：
Marraffini, Luciano A.
DOI：
10.1016/s1473-3099(15)00350-3
发表时间：
2016-04
期刊：
The Lancet. Infectious diseases
影响因子：
--
作者：
Kelly R;Zoubiane G;Walsh D;Ward R;Goossens H
通讯作者：
Goossens H
影响因子：
15.6
作者：
Malik, Danish J.;Sokolov, Ilya J.;Kirpichnikova, Anna
通讯作者：
Kirpichnikova, Anna
影响因子：
4.6
作者：
Krysiak-Baltyn, Konrad;Martin, Gregory J. O.;Gras, Sally L.
通讯作者：
Gras, Sally L.
DOI：
10.1111/nyas.12468
发表时间：
2014-01-01
期刊：
ANTIMICROBIAL THERAPEUTICS REVIEWS: INFECTIOUS DISEASES OF CURRENT AND EMERGING CONCERN
影响因子：
--
作者：
Allen, Heather K.;Trachsel, Julian;Casey, Thomas A.
通讯作者：
Casey, Thomas A.