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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Characterization of Coliphage PR772 and Evaluation of Its Use for Virus Filter Performance Testing
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Characterization of Coliphage PR772 and Evaluation of Its Use for Virus Filter Performance Testing

机译：Coliphage PR772的表征及其在病毒过滤器性能测试中的应用评估

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Virus filtration is a key clearance unit operation in the manufacture of recombinant protein, monoclonal antibody, and plasma-derived biopharmaceuticals. Recently, a consensus has developed among filter manufacturers and end users about the desirability of a common nomenclature and a standardized test for classifying and identifying virus-retentive filters. The Parenteral Drug Association virus filter task force has chosen PR772 as the model bacteriophage to standardize nomenclature for large-pore-size virus-retentive filters (filters designed to retain viruses larger than 50 to 60 nm in size). Previously, the coliphage PR772 (Tectiviridae family) has been used in some filtration studies as a surrogate for mammalian viruses of around 50 to 60 nm. In this report, we describe specific properties of PR772 critical to the support of its use for the standardization of virus filters. The complete genomic sequence of virulent phage PR772 was determined. Its genome contains 14,946 bp with an overall G+C content of 48.3 mol%, and 32 open reading frames of at least 40 codons. Comparison of the PR772 nucleotide sequence with the genome of Tectiviridae family prototype phage PRD1 revealed 97.2% identity at the DNA level. By dynamic light-scattering analysis, its hydrodynamic diameter was measured as 82 ± 6 nm, consistent with use in testing large-virus-retentive filters. Finally, dynamic light-scattering analysis of PR772 preparations purified on CsCl gradients showed that the phage preparations are largely monodispersed. In summary, PR772 appears to be an appropriate model bacteriophage for standardization of nomenclature for larger-pore-size virus-retentive filters.
机译：病毒过滤是重组蛋白，单克隆抗体和血浆衍生生物药物生产中关键的清除单元操作。最近，在过滤器制造商和最终用户之间已经达成共识，要求使用通用术语和用于分类和识别病毒保留过滤器的标准化测试。美国肠胃药物协会病毒过滤器工作组已选择PR772作为模型噬菌体，以标准化大孔径病毒保持过滤器（旨在保留尺寸大于50至60 nm的病毒的过滤器）的命名法。以前，大肠杆菌噬菌体PR772（Tectiviridae家族）已在一些过滤研究中用作替代大约50至60 nm的哺乳动物病毒的代用品。在此报告中，我们描述了PR772的特定属性，这些属性对于支持其用于病毒过滤器标准化至关重要。确定了毒性噬菌体PR772的完整基因组序列。它的基因组包含14946 bp，总G + C含量为48.3 mol％，并具有32个至少40个密码子的开放阅读框。 PR772核苷酸序列与Tectiviridae家族原型噬菌体PRD1的基因组进行比较，发现在DNA水平上的同源性为97.2％。通过动态光散射分析，测得其流体动力学直径为82±6 nm，与用于测试大型病毒保持过滤器的情况一致。最后，在CsCl梯度上纯化的PR772制剂的动态光散射分析表明，噬菌体制剂在很大程度上是单分散的。总之，PR772似乎是用于标准化大孔径病毒保持过滤器命名法的合适噬菌体。

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