Johnson Sarah, Brorson Kurt A, Frey Douglas D, Dhar Arun K, Cetlin David A
DBRRII, Office of Biotechnology Products, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, MD, 20993, USA.
Department of Chemical, Biochemical, and Environmental Engineering, University of Maryland Baltimore County, Baltimore, MD, 21250, USA.
Appl Biochem Biotechnol. 2017 Sep;183(1):318-331. doi: 10.1007/s12010-017-2447-y. Epub 2017 Mar 9.
Viral clearance is a critical aspect of biopharmaceutical manufacturing process validation. To determine the viral clearance efficacy of downstream chromatography and filtration steps, live viral "spiking" studies are conducted with model mammalian viruses such as minute virus of mice (MVM). However, due to biosafety considerations, spiking studies are costly and typically conducted in specialized facilities. In this work, we introduce the concept of utilizing a non-infectious MVM virus-like particle (MVM-VLP) as an economical surrogate for live MVM during process development and characterization. Through transmission electron microscopy, size exclusion chromatography with multi-angle light scattering, chromatofocusing, and a novel solute surface hydrophobicity assay, we examined and compared the size, surface charge, and hydrophobic properties of MVM and MVM-VLP. The results revealed that MVM and MVM-VLP exhibited nearly identical physicochemical properties, indicating the potential utility of MVM-VLP as an accurate and economical surrogate to live MVM during chromatography and filtration process development and characterization studies.
病毒清除是生物制药生产工艺验证的关键环节。为了确定下游色谱和过滤步骤的病毒清除效果，会使用诸如小鼠微小病毒（MVM）等模型哺乳动物病毒进行活病毒“加标”研究。然而，出于生物安全考虑，加标研究成本高昂，且通常在专门设施中进行。在这项工作中，我们引入了在工艺开发和表征过程中利用非感染性MVM病毒样颗粒（MVM-VLP）作为活MVM的经济替代物的概念。通过透射电子显微镜、多角度光散射尺寸排阻色谱、色谱聚焦和一种新型溶质表面疏水性测定法，我们检测并比较了MVM和MVM-VLP的大小、表面电荷和疏水性质。结果表明，MVM和MVM-VLP表现出几乎相同的物理化学性质，这表明MVM-VLP在色谱和过滤工艺开发及表征研究中作为活MVM的准确且经济的替代物具有潜在用途。
