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首页> 外文期刊>Applied Microbiology >Elution Is a Critical Step for Recovering Human Adenovirus 40 from Tap Water and Surface Water by Cross-Flow Ultrafiltration
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Elution Is a Critical Step for Recovering Human Adenovirus 40 from Tap Water and Surface Water by Cross-Flow Ultrafiltration

机译：洗脱是通过交叉流超滤从自来水和地表水中回收人腺病毒40的关键步骤

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This paper examines the recovery of the enteric adenovirus human adenovirus 40 (HAdV 40) by cross-flow ultrafiltration and interprets recovery values in terms of physicochemical interactions of virions during sample concentration. Prior to ultrafiltration, membranes were either blocked by exposure to calf serum (CS) or coated with a polyelectrolyte multilayer (PEM). HAdV 40 is a hydrophobic virus with a point of zero charge between pH 4.0 and pH 4.3. In accordance with predictions from the extended Derjaguin-Landau-Verwey-Overbeek theory, the preelution recovery of HAdV ( r _(pre)) from deionized water was higher with PEM-coated membranes ( r _(pre)~(PEM) = 74.8% ± 9.7%) than with CS-blocked membranes ( r _(pre)~(CS) = 54.1% ± 6.2%). With either membrane type, the total virion recovery after elution ( r _(post)) was high for both deionized water ( r _(post)~(PEM) = 99.5% ± 6.6% and r _(post)~(CS) = 98.8% ± 7.7%) and tap water ( r _(post)~(PEM) = 89% ± 15% and r _(post)~(CS) = 93.7% ± 6.9%). The nearly 100% recoveries suggest that the polyanion (sodium polyphosphate) and surfactant (Tween 80) in the eluent disrupt electrostatic and hydrophobic interactions between the virion and the membrane. Addition of EDTA to the eluent greatly improved the elution efficacy ( r _(post)~(CS) = 88.6% ± 4.3% and r _(post)~(PEM) = 87.0% ± 6.9%) with surface water, even when the organic carbon concentration in the water was high (9.4 ± 0.1 mg/liter). EDTA likely disrupts cation bridging between virions and particles in the feed water matrix or the fouling layer on the membrane surface. For complex water matrices, the eluent composition is the most important factor for achieving high virion recovery.IMPORTANCE Herein we present the results of a comprehensive physicochemical characterization of HAdV 40, an important human pathogen. The data on HAdV 40 surface properties enabled rigorous modeling to gain an understanding of the energetics of virion-virion and virion-filter interactions. Cross-flow filtration for concentration and recovery of HAdV 40 was evaluated, with postelution recoveries from ultrapure water (99%), tap water (～91%), and high-carbon-content surface water (～84%) being demonstrated. These results are significant because of the very low adenovirus recoveries that have been reported, to date, for other methods. The recovery data were interpreted in terms of specific interactions, and the eluent composition was designed accordingly to maximize HAdV 40 recovery.
机译：本文通过交叉流超滤检查了肠道腺病毒人腺病毒40（HAdV 40）的回收率，并根据样品浓缩过程中病毒体的物理化学相互作用解释了回收率值。在超滤之前，通过暴露于小牛血清（CS）封闭膜或用聚电解质多层膜（PEM）覆盖膜。 HAdV 40是一种疏水性病毒，在pH 4.0和pH 4.3之间具有零电荷点。根据扩展的Derjaguin-Landau-Verwey-Overbeek理论的预测，去离子水中HAdV（r _（pre））的预洗脱回收率较高，PEM涂层膜（r _（pre）〜（PEM）= 74.8） ％±9.7％）比CS阻滞膜（r_（pre）〜（CS）＝ 54.1％±6.2％）高。无论使用哪种膜，去离子水的洗脱后总病毒体回收率（r_（post））都很高（r_（post）〜（PEM）= 99.5％±6.6％和r_（post）〜（CS） = 98.8％±7.7％）和自来水（r_（post）〜（PEM）= 89％±15％和r_（post）〜（CS）= 93.7％±6.9％）。几乎100％的回收率表明洗脱液中的聚阴离子（聚磷酸钠）和表面活性剂（吐温80）破坏了病毒体与膜之间的静电和疏水相互作用。即使在以下情况下，用地表水添加EDTA也可以极大地提高洗脱效率（r_（post）〜（CS）= 88.6％±4.3％和r_（post）〜（PEM）= 87.0％±6.9％）。水中的有机碳浓度很高（9.4±0.1毫克/升）。 EDTA可能会破坏进水基质或膜表面污垢层中病毒体与颗粒之间的阳离子桥接。对于复杂的水基质，洗脱液组成是实现高病毒体回收率的最重要因素。重要事项本文中，我们提出了重要人类病原体HAdV 40的全面理化表征结果。 HAdV 40表面特性的数据可进行严格的建模，以了解病毒体-病毒体和病毒体-过滤器相互作用的能量。评估了错流过滤法对HAdV 40的浓缩和回收率，并证明了从超纯水（99％），自来水（〜91％）和高碳含量地表水（〜84％）中的洗脱后回收率。这些结果之所以重要，是因为迄今为止，对于其他方法，已经报道了非常低的腺病毒回收率。根据特定的相互作用解释了回收率数据，并相应设计了洗脱液成分，以最大限度地提高HAdV 40回收率。

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