Jannat Risat, Hsu David, Maheshwari Gargi
Fermentation and Cell Culture, Bioprocess R&D, Merck Research Laboratories, West Point, Pennsylvania 19486, USA.
Biotechnol Prog. 2005 Mar-Apr;21(2):446-50. doi: 10.1021/bp049812f.
Adenovirus shows significant promise as a vehicle for transfer of therapeutic genes into humans. Based on the importance of this viral vector, it is critical that adequate decontamination procedures are implemented during its large-scale production in multiproduct manufacturing facilities to prevent cross-product contamination and to reduce the risk of personnel exposure. Liquid decontamination procedures based on caustics are easily implemented in a manufacturing setting and are not corrosive to stainless steel surfaces at the concentrations found to inactivate viral proteins and nucleic acids. In this study, we have conducted small-scale experiments to determine the effectiveness of caustic inactivation procedures on adenovirus type 5 and have evaluated the robustness of the process to different sample matrices and adenovirus constructs. We find that the pH of a sample post-addition of caustic solution is a more accurate indicator of the effectiveness of the caustic than its concentration. We have demonstrated that a greater than 6 log reduction in the potency of adenovirus type 5 may be obtained upon exposure of the sample to sodium hydroxide and CIP-100 at concentrations greater than 0.09 M and 0.9%, respectively, at times greater than 10 min.
腺病毒作为将治疗性基因导入人体的载体显示出巨大的前景。基于这种病毒载体的重要性，在多产品生产设施中大规模生产腺病毒期间，实施适当的去污程序以防止交叉产品污染并降低人员接触风险至关重要。基于苛性碱的液体去污程序在生产环境中易于实施，并且在发现能使病毒蛋白和核酸失活的浓度下对不锈钢表面无腐蚀性。在本研究中，我们进行了小规模实验，以确定苛性碱灭活程序对5型腺病毒的有效性，并评估该过程对不同样品基质和腺病毒构建体的稳健性。我们发现，加入苛性碱溶液后样品的pH值比其浓度更能准确指示苛性碱的有效性。我们已经证明，当样品分别暴露于浓度大于0.09 M的氢氧化钠和0.9%的CIP-100中超过10分钟时，5型腺病毒的效力可降低大于6个对数。
