Michalik-Provasek Jordyn, Parker Harley, Lessor Lauren, Gill Jason J
Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, College Station, Texas, USA.
Center for Phage Technology, Texas A&M AgriLife Research and Texas A&M University, College Station, Texas, USA.
Phage (New Rochelle). 2021 Sep 1;2(3):112-119. doi: 10.1089/phage.2021.0005. Epub 2021 Sep 17.
Antimicrobial resistance in pathogenic bacteria is increasing worldwide. One solution to this crisis is bacteriophage therapy, a treatment that harnesses naturally occurring bacterial viruses to invade and lyse antimicrobial resistant bacterial hosts. In Gram-negative hosts, a by-product of bacteriophage production is bacterial endotoxin, which can cause serious immune reactions . Purification methods using organic solvent extraction can remove endotoxin in bacteriophage lysates. In this study, we investigate a method for removal of endotoxin from 16 high-titer lysates by extraction with 1-dodecanol, 1-octanol, dodecane, or decane. In these experiments, treatment with either 1-dodecanol or 1-octanol resulted in removal of 10-10 endotoxin units/mL. Recovery of bacteriophage in lysates treated with dodecanol without dialysis was >90%, and residual dodecanol was low (10-1500 ppm). Overall these results suggest that organic solvent extraction using 1-dodecanol is effective at removing bacterial endotoxin, maintaining bacteriophage titer, and reducing solvent contamination in 16 bacteriophage lysates.
全球范围内，致病细菌的抗微生物耐药性正在增强。解决这一危机的一种方法是噬菌体疗法，即利用天然存在的细菌病毒来侵入并裂解抗微生物耐药性细菌宿主的一种治疗方法。在革兰氏阴性宿主中，噬菌体产生的一种副产物是细菌内毒素，它可引发严重的免疫反应。使用有机溶剂萃取的纯化方法能够去除噬菌体裂解液中的内毒素。在本研究中，我们探究了一种用1-十二醇、1-辛醇、十二烷或癸烷萃取从16种高滴度裂解液中去除内毒素的方法。在这些实验中，用1-十二醇或1-辛醇处理可使内毒素单位/mL去除10-10。未经透析用十二醇处理的裂解液中噬菌体回收率>90%，且残留十二醇含量较低（10-1500 ppm）。总体而言，这些结果表明，使用1-十二醇进行有机溶剂萃取能有效去除细菌内毒素、维持噬菌体滴度并减少16种噬菌体裂解液中的溶剂污染。
