Stability and purity of a bacteriophage cocktail preparation for nebulizer delivery
Stability and purity of a bacteriophage cocktail preparation for nebulizer delivery
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DOI：
10.1111/lam.12161
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发表时间：
2014-02-01
影响因子：
2.4
通讯作者：
Maillard, J. -Y.
中科院分区：
文献类型：
--
作者：
Cooper, C. J.;Denyer, S. P.;Maillard, J. -Y.
本研究的目的是确定通过雾化递送的噬菌体混合物的稳定性和纯度。开发了一种对囊性纤维化患者的铜绿假单胞菌分离株具有活性的三种噬菌体混合物，用于潜在的雾化制剂。检查单个噬菌体随时间的活性保留，同时根据雾化产品的法规要求分析噬菌体混合物的细菌污染物和内毒素水平。使用连接到安德森级联冲击器的Porta-neb雾化器雾化噬菌体混合物。这三种酶在室温和4 ℃下储存180天后仍保持活性。在级联撞击采样器的较低阶段回收了雾化的空气，表明可能输送到肺部深处。噬菌体混合物符合细菌限度，但即使在纯化后，用鲎变形细胞溶解物（LAL）试验测量的内毒素水平仍大大超过可接受的水平。这些发现表明噬菌体的雾化是一种可行的递送选择;尽管需要适当的去热原策略来从基于噬菌体的制剂中去除细菌内毒素以满足法规要求。
The aim of this study was to determine the stability and purity of a phage cocktail to be delivered by nebulization. A cocktail of three phages active against Pseudomonas aeruginosa isolates from cystic fibrosis patients was developed for a potential nebulized formulation. The individual phages were examined for their retention of activity over time, while the phage cocktail was analysed for bacterial contaminant and endotoxin level according to regulatory requirements for nebulized products. The phage cocktail was nebulized using a Porta-neb nebulizer connected to an Anderson cascade impactor. The three phages retained activity over a period of 180days storage at room temperature and at 4 degrees C. Nebulized phages were recovered in the lower stages of the cascade impactor indicative of potential delivery deep into the lungs. The phage cocktail met bacterial limits but the endotoxin levels measured with the Limulus amoebocyte lysate (LAL) test remained considerably in excess of acceptable levels even following purification. These findings suggest that nebulization of phage is a viable delivery option; although, there is a need for appropriate depyrogenation strategies to remove bacterial endotoxins from phage-based preparations to meet regulatory requirements.