Development of a rapid method for the PEGylation of adenoviruses with enhanced transduction and improved stability under harsh storage conditions
Development of a rapid method for the PEGylation of adenoviruses with enhanced transduction and improved stability under harsh storage conditions
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DOI：
10.1089/10430340050111368
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发表时间：
2000-08-10
影响因子：
4.2
通讯作者：
Wilson, JM
中科院分区：
文献类型：
--
作者：
Croyle, MA;Yu, QC;Wilson, JM
聚乙二醇化是活化的单甲氧基聚乙二醇 (MPEG) 与治疗性蛋白质的游离赖氨酸基团的共价连接。这项技术增强了蛋白质的物理稳定性，并消除了针对它们产生的体液免疫反应。在本研究中，腺病毒载体用氰尿酰氯、琥珀酰亚胺基琥珀酸酯和三聚氯乙烷激活的 MPEG 进行修饰。在适当的缓冲条件下，反应在2小时内完成，聚乙二醇化腺病毒的转导效率不会因体外中和天然腺病毒抗体而受到影响。这些制剂在 42、25、4 和 -20 摄氏度储存后，保留的滴度显着高于未结合病毒。聚乙二醇化制剂在 -20 摄氏度下的稳定性表明，可以从制剂中消除甘油，而不会显着损失病毒滴度。通过气管内注射给药时，聚乙二醇化腺病毒在肺部的转导作用增加了两到三倍，在肝脏中的转导作用增加了五倍。静脉注射。
PEGylation is the covalent attachment of activated monomethoxy poly(ethylene) glycols (MPEGs) to free lysine groups of therapeutic proteins. This technology has enhanced the physical stability of proteins and ablated humoral immune responses generated against them. In this study, adenoviral vectors were modified with MPEGs activated by cyanuric chloride, succinimidyl succinate, and tresyl chloride. Under proper buffering conditions, reactions were complete within 2 hr, Transduction efficiency of PEGylated adenoviruses was not compromised by neutralizing antibodies to native adenovirus in vitro. These preparations retained titers that were significantly greater than those of the unconjugated virus after storage at 42, 25, 4, and -20 degrees C. Stability profiles of PEGylated preparations at -20 degrees C suggest that glycerol could be eliminated from formulations without significant loss of viral titer, PEGylated adenoviruses produced a two- to threefold increase in transduction in the lung when administered by intratracheal injection and a fivefold increase in transduction in the liver when administered intravenously.